N3 alkylated benzimidazole derivatives as mek inhibitors

ABSTRACT

Disclosed are amino acid prodrugs of compounds of the formula I  
                 
 
and pharmaceutically acceptable salts thereof, wherein W, R 1 , R 2 , R 7 , R 8 , R 9  and R 10  are as defined in the specification. Such compounds are MEK inhibitors and useful in the treatment of hyperproliferative diseases, such as cancer and inflammation, in mammals. Also disclosed is a method of using such compounds in the treatment of hyperproliferative disease in mammals, and pharmaceutical compositions containing such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Divisional application of U.S. patent applicationSer. No. 10/652,733 filed Aug. 29, 2003, which is a Continuation-in-Partapplication of U.S. patent application Ser. No. 10/387,879 filed Mar.13, 2003, which claims priority to U.S. Provisional Application Ser. No.60/364,007, filed Mar. 13, 2002, each of which is incorporated herein inits entirety by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to a series of alkylated(1H-Benzoimidazol-5-yl)-(4-substituted-phenyl)-amine derivatives thatare useful in the treatment of hyperproliferative diseases, such ascancer and inflammation, in mammals. This invention also relates to amethod of using such compounds in the treatment of hyperproliferativediseases in mammals, especially humans, and to pharmaceuticalcompositions containing such compounds.

2. Description of the State of the Art

Cell signaling through growth factor receptors and protein kinases is animportant regulator of cell growth, proliferation and differentiation.In normal cell growth, growth factors, through receptor activation (i.e.PDGF or EGF and others), activate MAP kinase pathways. One of the mostimportant and most well understood MAP kinase pathways involved innormal and uncontrolled cell growth is the Ras/Raf kinase pathway.Active GTP-bound Res results in the activation and indirectphosphorylation of Raf kinase. Raf then phosphorylates MEK1 and 2 on twoserine residues (S218 and S222 for MEK1 and S222 and S226 for MEK2) (Ahnet al., Methods in Enzymology 2001, 332, 417-431). Activated MEK thenphosphorylates its only known substrates, the MAP kinases, ERK1 and 2.ERK phosphorylation by MEK occurs on Y204 and T202 for ERK1 and Y185 andT183 for ERK2 (Ahn et al., Methods in Enzymology 2001, 332, 417-431).Phosphorylated ERK dimerizes and then translocates to the nucleus whereit accumulates (Khokhlatchev et al., Cell 1998, 93, 605-615). In thenucleus, ERK is involved in several important cellular functions,including but not limited to nuclear transport, signal transduction, DNArepair, nucleosome assembly and translocation, and mRNA processing andtranslation (Ahn et al., Molecular Cell 2000, 6, 1343-1354). Overall,treatment of cells with growth factors leads to the activation of ERK1and 2 which results in proliferation and, in some cases, differentiation(Lewis et al., Adv. Cancer Res. 1998, 74, 49-139).

In proliferative disease, genetic mutations and/or overexpression of thegrowth factor receptors, downstream signaling proteins, or proteinkinases involved in the ERK kinase pathway lead to uncontrolled cellproliferation and, eventually, tumor formation. For example, somecancers contain mutations which result in the continuous activation ofthis pathway due to continuous production of growth factors. Othermutations can lead to defects in the deactivation of the activatedGTP-bound Ras complex, again resulting in activation of the MAP kinasepathway. Mutated, oncogenic forms of Ras are found in 50% of colonand >90% pancreatic cancers as well as many others types of cancers(Kohl et al., Science 1993, 260, 1834-1837). Recently, bRaf mutationshave been identified in more than 60% of malignant melanoma (Davies, H.et al., Nature 2002, 417, 949-954). These mutations in bRaf result in aconstitutively active MAP kinase cascade. Studies of primary tumorsamples and cell lines have also shown constitutive or overactivation ofthe MAP kinase pathway in cancers of pancreas, colon, lung, ovary andkidney. (Hoshino, R. et al., Oncogene 1999, 18, 813-822). Hence, thereis a strong correlation between cancers and an overactive MAP kinasepathway resulting from genetic mutations.

As constitutive or overactivation of MAP kinase cascade plays a pivotalrole in cell proliferation and differentiation, inhibition of thispathway is believed to be beneficial in hyperproliferative disease. MEKis key player in this pathway as it downstream of Ras and Raf.Additionally, it is an attractive therapeutic target because the onlyknown substrates for MEK phosphorylation are the MAP kinases, ERK1 2.Inhibition of MEK has been shown to have potential therapeutic benefitin several studies. For example, small molecule MEK inhibitors have beenshown to inhibit human tumor growth in nude mouse xenagrafts,(Sebolt-Leopold et al., Nature-Medicine 1999, 5 (7), 810-816; Trachet etal., AACR Apr. 6-10, 2002, Poster #5426; Tecle, H. IBC 2^(nd)Inernational Conference of Protein Kinases, Sep. 9-10, 2002), blockstatic allodynia in animals (WO 01/05390 published Jan. 25, 2001) andinhibit growth of acute myeloid leukemia cells (Milella et al., J ClinInvest 2001, 108 (6), 851-859).

Small molecule inhibitors of MEK have been disclosed. At least thirteenpatent application have appeared in the last several years: U.S. Pat.No. 5,525,625 filed Jan. 24, 1995; WO 98/43960 published Oct. 8, 1998;WO 99/01421 published Jan. 14, 1999; WO 99/01426 published Jan. 14,1999; WO 00/41505 published Jul. 20, 2000; WO 00/42002 published Jul.20, 2000; WO 00/42003 published Jul. 20, 2000; WO 00/41994 publishedJul. 20, 2000; WO 00/42022 published Jul. 20, 2000; WO 00/42029published Jul. 20, 2000; WO 00/68201 published Nov. 16, 2000; WO01/68619 published Sep. 20, 2001; and WO 02/06213 published Jan. 24.2002.

SUMMARY OF THE INVENTION

This invention provides for alkylated(1H-benzoimidazol-5-yl)-(4-substituted phenyl)-amine compounds offormula I, and pharmaceutically acceptable salts and prodrugs thereofthat are useful in the treatment of hyperproliferative diseases.Specifically, the present invention relates to compounds of formula Ithat act as MEK inhibitors. Also provided is a method for treatment ofcancer. Also provided are formulations containing compounds of formula Iand methods of using the compounds to treat a patient in need thereof.In addition, there are described processes for preparing the inhibitorycompounds of formula I.

Accordingly, the present invention provides compounds of the formula I:

and pharmaceutically accepted salts, prodrugs and solvates thereof,wherein:

is an optional bond, provided that one and only one nitrogen of the ringis double-bonded;

R¹, R², R⁹ and R¹⁰ are independently selected from hydrogen, halogen,cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido,—OR³, —C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³, —NR⁴SO₂R⁶, —SO₂NR³R⁴,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —NR³R⁴, or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl), —S(O)_(j)(CR⁴R⁵)_(m)-aryl,aryl, arylalkyl, heteoraryl, heteroarylalkyl, heterocyclyl,heterocyclalkyl, —O(CR⁴R⁵)_(m)-aryl, —NR⁴(CR⁴R⁵)_(m)-aryl,—O(CR⁴R⁵)_(m)-heteroaryl, —NR⁴(CR⁴R⁵)_(m)-heteroaryl,—O(CR⁴R⁵)_(m)-heterocyclyl or —NR⁴(CR⁴R⁵)_(m)-heterocyclyl, where eachalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclylportion is optionally substituted with one to five groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R³ is selected from hydrogen, trifluoromethyl, C₁-C₁₀ alkyl, C₂-C₁₀alkenyl, C₂-C₁₀ alkynyl, (CH₂)_(n) C₃-C₁₀ cycloalkyl, C₃-C₁₀cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,heterocyclyl, heterocyclylalkyl, where each alkyl, alkenyl, alkynyl,cycloalkyl, aryl, heteroaryl and heterocyclyl portion optionallysubstituted with one to five groups independently selected from oxo,halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R″, —SO₂NR′R″, —C(O)R′, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″, —NR′C(O)R″, —C(O)NR′R″, —SR′, —S(O)R′, —SO₂R′,—NR′R″, —NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl,arylalkyl, heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R′, R″ and R′″ independently are selected from hydrogen, lower alkyl,lower alkenyl, aryl and arylalkyl;

R³ and R⁴ can be taken together with the atom to which they are attachedto form a 4 to 10 membered carbocyclic, heteroaryl or heterocyclic ring,each of which is optionally substituted with one to three groupsindependently selected from halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″, —SO₂NR′R″, —C(O)R′,—C(O)OR′, —OC(O)R′, —NR′C(O)OR′, —NR′C(O)R″, —C(O)NR′R″, —SO₂R′, —NR′R″,—NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R⁴ and R⁵ independently represent hydrogen or C₁-C₆ alkyl, or

R⁴ and R⁵ together with the atom to which they are attached form a 4 to10 membered carbocyclic, heteroaryl or heterocyclic ring, each of whichis optionally substituted with one to three groups independentlyselected from halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R″, —SO₂NR′R″, —C(O)R′, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″, —NR′C(O)R″, —C(O)NR′R″, —SO₂R′, —NR′R″,—NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R⁶ is selected from trifluoromethyl, C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl, where each alkyl, cycloalkyl, aryl, heteroaryl andheterocyclyl portion is optionally substituted with one to five groupsindependently selected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″, —SO₂NR′R″, —C(O)R′,—C(O)OR′, —OC(O)R′, —NR′C(O)OR″, —NR′C(O)R″, —C(O)NR′R″, —SO₂R′, —NR′R′,—NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R⁷ is selected from hydrogen, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, where eachalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclylportion is optionally substituted with one to five groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —SO₂R³, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

W is selected from heteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴,—C(O)NR⁴OR³, —C(O)R⁴OR³, —C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl),—C(O)(aryl), —C(O)(heteroaryl) and —C(O)(heterocyclyl), each of which isoptionally substituted with 1-5 groups independently selected from—NR³R⁴, —OR³, —R², or C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl,each of which is optionally substituted with 1 or 2 groups independentlyselected from —NR³R⁴ and —OR³;

R⁸ is selected from hydrogen, —SCF₃, —Cl, —Br, —F, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —OR³,—C(O)R³, —C(O)OR³, —NR⁴C(O)OR⁶, —OC(O)R³, —NR⁴SO₂R⁶, —SO₂NR³R⁴,—NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴, —NR³R⁴, or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl), —S(O)_(j)(CR⁴C⁵)_(m)-aryl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl, —O(CR⁴R⁵)_(m)-aryl, —NR₄(CR⁴R⁵)_(m)-aryl,—O(CR⁴R⁵)_(m)-heteroaryl, —NR⁴(CR⁴R⁵)_(m)-heteroaryl,—O(CR⁴R⁵)_(m)-heterocyclyl or —NR⁴(CR⁴R⁵)_(m)-heterocyclyl, where eachalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclylportion is optionally substituted with one to five groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

m is 0, 1, 2, 3, 4 or 5; and

j is 1 or 2.

DETAILED DESCRIPTION OF THE INVENTION

The novel compounds encompassed by the instant invention are thosedescribed by the general formula I set forth above, and thepharmaceutically acceptable salts and prodrugs thereof.

The present invention also provides compounds of formula I in which R⁷is C₁-C₁₀ alkyl, C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, C₃-C₇heterocycloalkyl or C₃-C₇ heterocycloalkylalkyl each of which can beoptionally substituted with 1-3 groups independently selected from oxo,halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³,—OC(O)R³, —SO₂R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl.

The present invention also provides compounds of formula I wherein R⁸ is—OCF₃, Br or Cl, R² is hydrogen, and R¹ is lower alkyl or halogen.

The present invention also provides compounds of formula I wherein R⁹ ishydrogen or halogen, and R¹⁰ is halogen.

The present invention also provides compounds of formula I wherein W is—C(O)OR³ or —C(O)NR⁴OR³.

The present invention also provides compounds of formula II:

wherein W, R¹, R⁷, R⁸, R⁹ and R¹⁰ are as defined above for formula I.

The present invention also provides compounds of formula II in which R⁷is C₁-C₁₀ alkyl, C₃-C₇ cycloalkyl or C₃-C₇ cycloalkyl, each of which canbe optionally substituted with 1-3 groups independently selected fromoxo, halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³,—OC(O)R³, —SO₂R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl.

The present invention also provides compounds of formula II wherein R⁸is —OCF₃, Br or Cl, and R¹ is lower alkyl or halogen.

The present invention also provides compounds of formula II wherein R⁹is hydrogen or halogen, and R¹⁰ is hydrogen.

The present invention also provides compounds of formula II wherein W is—C(O)OR³ or —C(O)NR⁴OR³.

The present invention also provides compounds of formula III:

wherein R¹, R², R⁷, R⁸ and R⁹ are as defined above for formula I, and Ais —OR³ or —NR⁴OR³, wherein R³ and R⁴ are as defined above for formulaI.

The present invention also provides compounds of formula III in which R⁷is C₁-C₁₀ alkyl, C₃-C₇ cycloalkyl or C₃-C₇ cycloalkylalkyl, each ofwhich can be optionally substituted with 1-3 groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —SO₂R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl.

The present invention also provides compounds of formula III wherein R⁸is —OCF₃, Br or Cl, and R² is hydrogen, and R¹ is lower alkyl orhalogen.

The present invention also provides compounds of formula III wherein R⁹is hydrogen or halogen.

The present invention also provides compounds of formula III wherein R³is hydrogen or lower alkyl when A is —OR³, and R⁴ is hydrogen when A is—NR⁴OR³.

The present invention also provides compounds of formula IIIa:

wherein R¹, R², R⁷, R⁸ and R⁹ are as defined above for formula I, and Ais —OR³ or —NR⁴OR³, wherein R³ and R⁴ are as defined above for formulaI.

The present invention also provides compounds of formula IIIa in whichR⁷ is C₁-C₁₀ alkyl, C₃-C₇ cycloalkyl or C₃-C₇ cycloalkylalkyl, each ofwhich can be optionally substituted with 1-3 groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —SO₂R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl.

The present invention also provides compounds of formula IIIa wherein R⁸is —OCF₃, Br or Cl, R² is hydrogen, and R¹ is lower alkyl or halogen.

The present invention also provides compounds of formula IIIa wherein R⁹is hydrogen or halogen.

The present invention also provides compounds of formula IIIa wherein R³is hydrogen or lower alkyl when A is —OR³, and R⁴ is hydrogen when A is—NR⁴OR³.

The present invention also provides compounds of formula IIIb:

wherein R¹, R⁷, R⁸ and R⁹ are defined above for formula I, and A is —OR³or —NR⁴OR³, wherein R³ and R⁴ are as defined above for formula I.

The present invention also provides compounds of formula IIIb in whichR⁷ is C₁-C₁₀ alkyl, C₃-C₇ cycloalkyl or C₃-C₇ cycloalkylalkyl, each ofwhich can be optionally substituted with 1-3 groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —SO₂R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl.

The present invention also provides compounds of formula IIIa wherein R⁸is —OCF₃, Br or Cl, and R¹ is lower alkyl or halogen.

The present invention also provides compounds of formula III wherein R⁹is fluoro or chloro.

The present invention also provides compounds of formula III wherein R³is hydrogen or lower alkyl when A is —OR³; and R⁴ is hydrogen when A is—NR⁴OR³.

Except as expressly defined otherwise, the following definition of termsis employed throughout this specification.

By “C₁-C₁₀ alkyl”, “alkyl” and “lower alkyl” in the present invention ismeant straight or branched chain alkyl groups having 1-10 carbon atoms,such as, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl,tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl,3-hexyl, 3-methylpentyl, heptyl, octyl, and the like. Preferred alkylradicals are C₁₋₆ alkyl. More preferred alkyl radicals are C₁₋₃ alkyl.

By “C₂-C₁₀ alkenyl”, “lower alkenyl” and “alkenyl” means straight andbranched hydrocarbon radicals having from 2 to 10 carbon atoms and atleast one double bond and includes ethenyl, propenyl, 1-but-3-enyl,1-pent-3-enyl, 1-hex-5-enyl and the like. More preferred are loweralkenyl having 3-5 carbon atoms.

By “C₂-C₁₀ alkynyl”, “lower alkynyl” and “alkynyl” means straight andbranched hydrocarbon radicals having from 2 to 10 carbon atoms and atleast one triple bond and includes ethynyl, propynyl, butynyl,pentyn-2-yl and the like. More preferred are alkynyl having 3-5 carbonatoms.

By the term “halogen” in the present invention is meant fluorine,bromine, chlorine, and iodine.

By “aryl” is meant an aromatic carbocylic group having a single ring(e.g., phenyl), multiple rings (e.g., biphenyl), or multiple condensedrings in which at least one is aromatic, (e.g.,1,2,3,4-tetrahydronaphthyl, naphthyl), which is optionally mono-, di-,or trisubstituted with, e.g., halogen, lower alkyl, lower alkoxy,trifluoromethyl, aryl, heteroaryl, and hydroxy.

By “heteroaryl” is meant one or more aromatic ring systems of 5-, 6-, or7-membered rings which includes fused ring systems (at least one ofwhich is aromatic) of 5-10 atoms containing at least one and up to fourheteroatoms selected from nitrogen, oxygen, or sulfur. Examples ofheteroaryl, groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl,triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl,benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl,phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl,oxadiazolyl, triazolyl, thiadiazolyl, thiadiazolyl, furazanyl,benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl,quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Spiromoieties are also included within the scope of this definition.Heteroaryl groups are optionally mono-, di-, or trisubstituted with,e.g., halogen, lower alkyl, lower alkoxy, haloalkyl, aryl, heteroaryl,and hydroxy.

As used herein, the term “carbocycle”, “carbocyclyl”, “cycloalkyl” or“C₃-C₁₀ cycloalkyl” refers to saturated carbocyclic radicals havingthree to ten carbon atoms. The cycloalkyl can be monocyclic, or apolycyclic fused system, and can be fused to an aromatic ring. Examplesof such radicals include cyclopropyl, cyclobutyl, cyclopentyl,cyclopentyl and cyclohexyl. The cycloalkyl groups herein areunsubstituted or, as specified, substituted in one or more substitutedpositions with various groups. For example, such cycloalkyl groups maybe optionally substituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy,halogen, hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkyl ordi(C₁-C₆)alkylamino(C₁-C₆)alkyl.

By “heterocycle” or “heterocyclyl” is meant one or more carbocyclic ringsystem of 5-, 6-, or 7-membered rings rings which includes fused ringsystems of 4-10 atoms containing at least one and up to four heteroatomsselected from nitrogen, oxygen, or sulfur, and with the proviso that thering of the group does not contain two adjacent O or S atoms. A fusedsystem can be a heterocycle fused to an aromatic group. Preferredheterocycles include, but are not limited to, pyrrolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl,oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl,diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridynyl, 2-pyrrolinyl,3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl,1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl,dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl,imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,azabicyclo[2.2.2]hexanyl, 3H-indolyl and quinolizinyl. Spiro moietiesare also included within the scope of the definition. The foregoinggroups, as derived from the groups listed above, may be C-attached orN-attached where such is possible. For instance, a group derived frompyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached).Further, a group derived from imidazole may be imidazol-1-yl(N-attached) or imidazol-3-yl (C-attached). An example of a heterocyclicgroup wherein 2 ring carbon atoms are substituted with oxo (═O) moietiesis 1,1-dioxo-thiomorpholinyl. The heterocycle groups herein areunsubstituted or, as specified, substituted in one or more substitutedposition with various groups. For example, such heterocycle groups maybe optionally substituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy,hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkyl ordi(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The term “arylalkyl” means an alkyl moiety (as defined above)substituted with one or more aryl moiety (also as defined above). Morepreferred arylalkyl radicals are aryl-C₁₋₃-alkyls. Examples includesinclude benzyl, and the like.

The term “heteroarylalkyl” means an alkyl moiety (as defined above)substituted with a heteroaryl moiety (also as defined above). Morepreferred heteroarylalkyl radicals are 5- or 6-memberedheteroaryl-C₁₋₃-alkyls. Examples include oxazolylmethyl, pyridylethyland the like.

The term “heterocyclylalkyl” means an alkyl moiety (as defined above)substituted with a heterocyclyl moiety (also defined above). Morepreferred heterocyclylalkyl radicals are 5- or 6-memberedheterocyclyl-C₁₋₃-alkyls. Examples include tetrahydropyranylmethyl.

The term “cycloalkylalkyl” means an alkyl moiety (as defined above)substituted with a cycloalkyl moiety (also defined above). Morepreferred heterocyclyl radicals are 5- or 6-memberedcycloalkyl-C₁₋₃-alkyls. Examples include cyclopropylmethyl.

The term “Me” means methyl, “Et” means ethyl, “Bu” means butyl and “Ac”means acetyl.

The phrase “pharmaceutically acceptable salt(s)”, as used herein, unlessotherwise indicated, includes salts of acidic and basic groups which maybe present in the compounds of the present invention. The compounds ofthe present invention that are basic in nature are capable of forming awide variety of salts with various inorganic and organic acids. Theacids that may be used to prepare pharmaceutically acceptable acidaddition salts of such basic compounds of the present invention arethose that form non-toxic acid addition salts, i.e., salts containingpharmaceutically acceptable anions, such as the acetate,benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate,bromide, calcium, camsylate, carbonate, chloride, clavulanate, citrate,dihydrochloride, edislyate, estolate, esylate, ethylsuccinate, fumarate,gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate,hydrabamine, hydrobromide, hydrochloride, iodide, isothionate, lactate,lactobionate, laurate, malate, maleate, mandelate, mesylate,methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamoate(embonate), palimitate, pantothenate, phospate/diphosphate,polygalacturonate, salicylate, stearate, subacetate, succinate, tannate,tartrate, teoclate, tosylate, triethiodode, and valerate salts. Since asingle compound of the present invention may include more than oneacidic or basic moieties, the compounds of the present invention mayinclude mono, di or tri-salts in a single compound.

In the case of an acidic moiety in compound of the present invention, asalt may be formed by treatment of a compound of the present inventionwith a basic compound, particularly an inorganic base. Preferredinorganic salts are those formed with alkali and alkaline earth metalssuch as lithium, sodium, potassium, barium and calcium. Preferredorganic base salts include, for example, ammonium, dibenzylammonium,benzylammonium, 2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium,phenylethylbenzylamine, dibenzyl-ethylenediamine, and the like salts.Other salts of acidic moieties may include, for example, those saltsformed with procaine, quinine and N-methylglusoamine, plus salts formedwith basic amino acids such as glycine, omithine, histidine,phenylglycine, lysine and arginine. An especially preferred salt is asodium or potassium salt of a compound of the present invention.

With respect to basic moieties, a salt is formed by the treatment of acompound of the present invention with an acidic compound, particularlyan inorganic acid. Preferred inorganic salts of this type may include,for example, the hydrochloric, hydrobromic, hydroiodic, sulfuric,phosphoric or the like salts. Preferred organic salts of this type, mayinclude, for example, salts formed with formic, acetic succinic, citric,lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic, D-glutamic,D-camphoric, glutaric, glycolic, phthalic, tartaric, stearic,salicyclic, methanesulfonic, benzenesulfonic, paratoluenesulfonic,sorbic, puric, benzoic, cinnamic and the like organic acids. Anespecially preferred salt of this type is a hydrochloride or sulfatesalt of a compound of the present invention.

In the compounds of the present invention, where terms such as(CR⁴R⁵)_(m) or (CR⁴R⁵)_(t) are used, R⁴ and R⁵ may vary each iterationof m or t above 1. For instance, where m or t is 2, the terms(CR⁴R⁵)_(m) or (CR⁴R⁵)_(t) may equal —CH₂CH₂— or—CH(CH₃)C(CH₂CH₃)(CH₂CH₂CH₃)— or any number of similar moieties fallingwithin the scope of the definitions of R⁴ and R⁵.

Certain compounds of the present invention may have asymmetric centersand therefore exist in different enantiomeric forms. All optical isomersand stereoisomers of the compounds of the present invention, andmixtures thereof, are considered to be within the scope of theinvention. With respect to the compounds of the present invention, theinvention includes the use of a racemate, one or more enantiomericforms, one or more diastereomeric forms, or mixtures thereof. Thecompounds of the present invention may also exist as tautomers. Thisinvention relates to the use of all such tautomers and mixtures thereof.

The subject invention also includes isotopically-labeled compounds,which are identical to those recited in the present invention, but forthe fact that one or more atoms are replaced by an atom having an atomicmass or mass number different from the atomic mass or mass numberusually found in nature. Examples of isotopes that can be incorporatedinto compounds of the invention include isotopes of hydrogen, carbon,nitrogen, oxygen, phosphorous, sulfur, fluorine and chloride, such as²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl,respectively. Compounds of the present invention prodrugs thereof, andpharmaceutically acceptable salts of said compounds or of said prodrugswhich contain the aforementioned isotopes and/or other isotopes of otheratoms are within the scope of this invention. Certainisotopically-labeled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H and ¹⁴C areincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, i.e., ³H and carbon-14, i.e., ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.Further, substitution with heavier isotopes such as deuterium, i.e., ²H,can afford certain therapeutic advantages resulting from greatermetabolic stability, for example increased in vivo half-life or reduceddosage requirements and, hence may be preferred in some circumstances.Isotopically labeled compound of the present invention and prodrugsthereof cam generally be prepared by carrying out procedures disclosedin the Schemes and/or in the Examples and Preparations below, bysubstituting a readily available isotopically reagent for anon-isotopically labeled reagent.

This invention also encompasses pharmaceutical compositions containing aprodrugs of compound of Formula I-IIIb, and methods of treatingproliferative disorders, or abnormal cell growth by administeringprodrugs of compounds of the present invention. Compounds of the presentinvention having free amino, amido, hydroxy or carboxylic groups can beconverted into prodrugs. Prodrugs include compounds wherein an aminoacid residue, or a polypeptide chain of two or more (e.g., two, three orfour) amino acid residues is covalently joined through an amide or esterbond to a free amino, hydroxy or carboxylic acid group of compounds ofthe present invention. The amino acid residues include but are notlimited to the 20 naturally occurring amino acids commonly designated bythree letter symbols and also includes 4-hydroxyproline, hydroxylysine,demosine, isodemosine, 3-methylhistidine, noryaline, beta-alanine,gamma-aminobutyric acid, cirtulline, homocysteine, homoserine, omithineand methionine sulfone. One preferred prodrug comprises a compound ofthis invention covalently coupled to a valine residue. Additional typesof prodrugs are also encompassed. For instance, free carboxyl groups canbe derivatized as amides or alkyl esters. Free hydroxy groups may bederivatized using groups including but not limiting to hemisuccinates,phosphate, esters, dimethylaminoacetates, andphosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug DeliveryReviews 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate ester andsulfate ester of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including but notlimited to ether, amino and carboxylic acid functionalities, or wherethe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med. Chem. 1996,39, 10. Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding but not limited to ether, amino and carboxylic acidfunctionalities.

It is to be understood that in instances where two or more radicals areused in succession to define a substituent attached to a structure, thefirst named radical is considered to be terminal and the last namedradical is considered to be attached to the structure in question. Thus,for example, the radical arylalkyl is attached to the structure inquestion by the alkyl group.

The invention also relates to a pharmaceutical composition for thetreatment of a hyperproliferative disorder in a mammal which comprises atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof, and apharmaceutically acceptable carrier. In one embodiment, saidpharmaceutical composition is for the treatment of cancer such as brain,lung, squamous cell, bladder, gastric, pancreatic, breast, head, neck,renal, kidney, ovarian, prostate, colorectal, esophageal, testicular,gynecological or thyroid cancer. In another embodiment, saidpharmaceutical composition is for the treatment of a non-canceroushyperproliferative disorder such as benign hyperplasia of the skin(e.g., psoriasis), restenosis, or prostate (e.g., benign prostatichypertrophy (BPH)).

The invention also relates to a pharmaceutical composition for thetreatment of pancreatitis or kidney disease (including proliferativeglomerulonephritis and diabetes-induced renal disease) or the treatmentof pain in a mammal which comprises a therapeutically effective amountof a compound of the present invention, or a pharmaceutically acceptablesalt, prodrug or hydrate thereof, and a pharmaceutically acceptablecarrier.

The invention also relates to a pharmaceutical composition for theprevention of blastocyte implantation in a mammal which comprises atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof, and apharmaceutically acceptable carrier.

The invention also relates to a pharmaceutical composition for treatinga disease related to vasculogenesis or angiogenesis in a mammal whichcomprises a therapeutically effective amount of a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug orhydrate thereof, and a pharmaceutically acceptable carrier. In oneembodiment, said pharmaceutical composition is for treating a diseaseselected from the group consisting of tumor angiogenesis, chronicinflammatory disease such as rheumatoid arthritis, atherosclerosis,inflammatory bowel disease, skin disease such as psoriasis, eczema, andscleroderma, diabetes, diabetic retinopathy, retinopathy of prematurity,age-related macular degeneration, hemangioma, glioma, melanoma, Kaposi'ssarcoma and ovarian, breast, lung, pancreatic, prostate, colon andepidermoid cancer.

The invention also relates to a method of treating a hyperproliferativedisorder in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof. Inone embodiment, said method relates to the treatment of cancer such asbrain, lung, squamous cell, bladder, gastric, pancreatic, breast, head,neck, renal, kidney, ovarian, prostate, colorectal, esophageal,testicular, gynecological or thyroid cancer. In another embodiment, saidmethod relates to the treatment of a non-cancerous hyperproliferativedisorder such as benign hyperplasia of the skin (e.g., psoriasis),restenosis, or prostate (e.g., benign prostatic hypertrophy (BPH)).

The invention also relates to a method for the treatment of ahyperproliferative disorder in a mammal that comprises administering tosaid mammal a therapeutically effective amount of a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug orhydrate thereof, in combination with an anti-tumor agent selected fromthe group consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, growth factor inhibitors,cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors,biological modifiers, anti-hormones, angiogenesis inhibitors, andanti-androgens.

The invention also relates to a method of treating pancreatitis orkidney disease in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof.

The invention also relates to a method of preventing blastocyteimplantation in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof.

The invention also relates to a method of treating disease related tovasculogenesis or angiogenesis in a mammal which comprises administeringto said mammal a therapeutically effective amount of a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug orhydrate thereof. In one embodiment, said method is for treating adisease selected from the group consisting of tumor angiogenesis,chronic inflammatory disease such as rheumatoid arthritis,atherosclerosis, inflammatory bowel disease, skin disease such aspsoriasis, eczema, and scleroderma, diabetes, diabetic retinopathy,retinopathy of prematurity, age-related macular degeneration,hemangioma, glioma, melanoma, Kaposi's sarcoma and ovarian, breast,lung, pancreatic, prostate, colon and epidermoid cancer.

Patients that can be treated with compounds of the present invention, orpharmaceutically acceptable salts, prodrugs and hydrates of saidcompounds, according to the methods of this invention include, forexample, patients that have been diagnosed as having psoriasis,restenosis, atherosclerosis, BPH, long cancer, bone cancer, CMML,pancreatic cancer, skin cancer, cancer of the head and neck, cutaneousor intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer,cancer of the anal region, stomach cancer, colon cancer, breast cancer,testicular, gynecologic tumor (e.g., uterine sarcomas, carcinoma of thefallopian tubes, carcinoma of the endometrium, carcinoma of the cervix,carcinoma of the vagina or carcinoma of the vulva), Hodgkin's disease,cancer of the esophagus, cancer of the small intestine, cancer of theendocrine system (e.g., cancer of the thyroid, parathyroid or adrenalglands), sarcomas of soft tissue, cancer of the urethra, cancer of thepenis, prostate cancer, chronic or acute leukemia, solid tumor ofchildhood, lymphocytic lymphomas, cancer of the bladder, cancer of thekidney or ureter (e.g., renal cell carcinoma, carcinoma of the renalpelvis), or neoplasms of the central nervous system (e.g., primary CNSlymphoma, spinal axis tumors, brain stem gliomas or pituitary adenomas).

This invention also relates to a pharmaceutical composition forinhibiting abnormal cell growth in a mammal which comprises an amount ofa compound of the present invention, or a pharmaceutically acceptablesalt or solvate or prodrug thereof, in combination with an amount of achemotherapeutic, wherein the amounts of the compound, salt, solvate, orprodrug, and of the chemotherapeutic are together effective ininhibiting abnormal cell growth. Many chemotherapeutics are presentlyknown in the art. In one embodiment, the chemotherapeutic is selectedfrom the group consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, growth factor inhibitors,cell cycle inhibitors, enzymes, topoisomerase inhibitors, biologicalresponse modifiers, anti-hormones, angiogenesis inhibitors, andanti-androgens.

This invention further relates to a method for inhibiting abnormal cellgrowth in a mammal or treating a hyperproliferative disorder whichmethod comprises administering to the mammal an amount of a compound ofthe present invention, or a pharmaceutically acceptable salt or solvateor prodrug thereof, in combination with radiation therapy, wherein theamount of the compound, salt, solvate, or prodrug, is in combinationwith the radiation therapy effective in inhibiting abnormal cell growthor treating the hyperproliferative disorder in the mammal. Techniquesfor administering radiation therapy are known in the art, and thesetechniques can be used in the combination therapy described herein. Theadministration of the compound of the invention in this combinationtherapy can be determined as described herein.

It is believed that the compounds of the present invention can renderabnormal cells more sensitive to treatment with radiation for purposesof killing and/or inhibiting the growth of such cells. Accordingly, thisinvention further relates to a method for sensitizing abnormal cells ina mammal to treatment with radiation which comprises administering tothe mammal an amount of a compound of the present invention orpharmaceutically acceptable salt or solvate or prodrug thereof, whichamount is effective is sensitizing abnormal cells to treatment withradiation. The amount of the compound, salt, or solvate in this methodcan be determined according to the means for ascertaining affectiveamount of such compounds described herein.

The invention also relates to a method of and to a pharmaceuticalcomposition of inhibiting abnormal cell growth in a mammal whichcomprises an amount of a compound of the present invention, or apharmaceutically acceptable salt or solvate thereof, a prodrug thereof,or an isotopically-labeled derivative thereof, and an amount of one ormore substances selected from anti-angiogenesis agents, signaltransduction inhibitors, and antiproliferative agents.

Anti-angiogenesis agents, such as MMP-2 (matrix-metalloprotienase 2)inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and COX-II(cyclooxygenase II) inhibitors, can be used in conjunction with acompound of the present invention and pharmaceutical compositiondescribed herein. Examples of useful COX-II inhibitors include CELEBREX™(alecoxib), valdecoxib, and rofecoxib. Examples of useful matrixmetalloprotienase inhibitors are described in WO 96/33172 (publishedOct. 24, 1996), WO 96/27583 (published Mar. 7, 1996). European PatentApplication No. 97304971.1 (filed Jul. 8, 1997), European PatentApplication No. 99308617.2 (filed Oct. 29, 1999), WO 98/07697 (publishedFeb. 26, 1998), WO 98/03516 (published Jan. 29, 1998), WO 98/34918(published Aug. 13, 1998), WO 98/34915 (published Aug. 13, 1998), WO98/33768 (published Aug. 6, 1998), WO 98/30566 (published Jul. 16,1998), European Patent Publication 606,046 (published Jul. 13, 1994),European Patent Publication 931,788 (published Jul. 28, 1999), WO90/05719 (published May 31, 1990), WO 99/52910 (published Oct. 21,1999), WO 99/52889 (published Oct. 21, 1999), WO 99/29667 (publishedJun. 17, 1999), PCT International Application No. PCT/IB98/01113 (filedJul. 21, 1998), European Patent Application No. 99302232.1 (filed Mar.25, 1999), Great Britain Patent Application No. 9912961.1 (filed Jun. 3,1999), U.S. Provisional Application No. 60/148,464 (filed Aug. 12,1999), U.S. Pat. No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No.5,861,510 (issued Jan. 19, 1999), and European Patent Publication780,386 (published Jun. 25, 1997), all of which are incorporated hereinin their entireties by reference. Preferred MMP-2 and MMP-9 inhibitorsare those that have little or no activity inhibiting MMP-1. Morepreferred, are those that selectively inhibit MMP-2 and/or MMP-9relative of the other matrix-metalloproteinases (i.e., MMP-1, MMP-3,MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Some specific examples of MMP inhibitors useful in the present inventionare AG-3340, RO 32-3555, and RS 13-0830.

The terms “abnormal cell growth” and “hyperproliferative disorder” areused interchangeably in this application.

“Abnormal cell growth”, as used herein, unless otherwise indicated,refers to cell growth that is independent of normal regulatorymechanisms (e.g., loss of contact inhibition). This includes, forexample, the abnormal growth of: (1) tumor cells (tumors) thatproliferate by expressing a mutated tyrosine kinase or overexpression ofa receptor tyrosine kinase; (2) benign and malignant cells of otherproliferative disease in which aberrant tyrosine kinase activationoccurs; (3) any tumors that proliferate by receptor tyrosine kinases;(4) any tumors that proliferate by aberrant serine/threonine kinaseactivation; and (5) benign and malignant cells of other proliferativedisease in which aberrant serine/threonine kinase activation occurs.

The term “treating”, as used herein, unless otherwise indicated, meansreversing, alleviating, inhibiting the process of, or preventing thedisorder or condition to which such term applies, or one or moresymptoms of such disorder or condition. The term “treatment,” is usedherein, unless otherwise indicated, refers to the act of treating as“treating” is defined immediately above.

Representative compounds of the present invention, which are encompassedby the present invention include, but not limited to the compounds ofthe examples and their pharmaceutically acceptable acid or base additionsalts or prodrugs thereof. General synthetic methods which may bereferred to for preparing some of the compounds of the present inventionare provided in PCT published application number WO 00/42022 (publishedJul. 20, 2000). The foregoing patent application is incorporated hereinby reference in its entirety. Illustrations of several methods for thepreparation of compounds of the present invention are shown in Schemes1-4.

Scheme 1 illustrates the synthesis of compounds of the presentinvention. In step 1, the acid is nitrated using standard conditionspreferable fuming nitric acid H₂SO₄. In step 2, the aniline is preparedby fluoride displacement with NH₄OH at room temperature in waterfollowed by careful acidification with concentrated mineral acid to pHnear 0. In step 3, the ester is prepared by standard methods includingby not limited to Fisher Esterification (MeOH, H₂SO₄), and reaction withTMSCHN₂ in suitable organic solvents like PhMe/MeOH or THF/MeOH. In step4, the dianilino derivative is prepared by heating (60 to 200°C.) theester with an excess of the appropriate aniline neat or in an organicsolvent like xylenes. For example, when R¹=Me and R²=H the preferredmethod is stirring the ester with 10 equivalents aniline in xylenes atreflux until complete reaction. In step 5, the nitro arene is reduced toproduce the diamine by standard reduction conditions, including by notlimited to H₂, and Pd/C or Pd(OH)₂/C or Raney Nickel in organic solventlike EtOH or THF. Fe in AcOH, Zn in AcOH or Zn, NH₄Cl (aq) in MeOH. Instep 6, the diamine is cyclization by heating with formic acid neat orformamidine acetate in an appropriate solvent like EtOH. Alternatively,when R¹or R² does not equal halo the nitro arene can be converteddirectly to the benzimidazole in step 7 by heating in formic acid withPd(OH)₂/C or other palladium source like Pd/C. In step 8, a halide canbe incorporated by standard methods, including but not limited to NBS orNCS and pTsOH in organic cosolvents like THF and MeOH. In step 9, thebenzimidazole is alkylated to give a near equal mixture of N1 and N3products which are separable by standard techniques, including, forexample, chromatography and trituration. The alkylation is accomplishedby use of an alkylating agent like an alkyl halide and base like NaH, orK₂CO₃ in suitable organic solvent like DMF or THF at temperature rangingfrom 0 to 80° C. R⁷ can be further modified by various synthetic methodsknown in the art, as exemplified below. In step 10, the ester ishydrolysized by standard saponification methods. The acids is thenconverted to the desired hydroxamate in step 11 by standard couplingprocedures including but not limited to EDCl, HOBt or PyBOP and theappropriate hydroxylamine in suitable organic solvents like DMF, THF ormethylene chloride.

Scheme 2 illustrates an example in which the R⁸ substituent is on theaniline prior to the coupling procedure with the nitro ester. Thereaction description is exactly like that for Scheme 1 except that thereis no need to incorporated R⁸ as it is present in the aniline from thebeginning.

In Scheme 3, the preparation of N3 alkyl amino benzimidazole derivativesis illustrated. In step 1, the terminal alkene of the N3 alkylatedbenzimidazole hydroxamate is dihydroxylated using a suitable oxidantlike OsO₄ in suitable solvent or KMnO₄ or I₂, AgOAc, AcOH, water. Thediol is then further oxidized in step 2 by NaIO₄ or Pb(OAc)₄ in suitablebiphasic mixture to give the aldehyde. Alternatively (step 3), thealkene can be directly converted to the aldehyde by standard methodsincluding but not limited to ozone/Me₂S, NaIO₄/OsO₄ or KMnO₄. In step 4,the amine is prepared by reductive amination using standard methods suchas Na(CN)BH₃, Na(OAc)₃BH, NMe₄BH(OAc)₃ with or without AcOH in asuitable solvent such as methylene chloride, acetonitrile or THF. Thepreferable reduction amination is to treat the aldehyde with amine,Me₄NBH(OAc)₃ and acetic acid in MeCN at room temperature.

Scheme 4 illustrates the preparation of compounds of the presentinvention where W is heterocyclic. In step 1, the methyl ester isconverted to the hydrazide by stirring with hydrazine in a suitablesolvent like EtOH at temperatures from 50 to 100° C. The desiredheterocyclic derivative is then prepared by cyclization with theappropriate reagent. For oxadiazole 21 the hydrazide is treated with anorthoformate like triethyl orthoformate, and an acid catalyst like pTsOHin a suitable organic solvent like EtOH at elevated temperatures(50-100° C.). For hydroxy oxadiazole 22 the hydrazide can be cyclizedwith phosgene or a phosgene equivalent like triphosgene or carbonyldiimidazole in a suitable organic solvent like toluene at temperaturesranging from 50 to 120° C. The mercapto oxadizaole 23 can be prepared byreaction with carbon disulfide, and base like KOH in suitable organicsolvent like EtOH at elevated temperatures (50-100° C.). The aminooxadiazole 24 can be made by reaction with BrCN and base like NaHCO₃, ina suitable biphasic solvent system like dioxane and water at roomtemperature. Finally, the substituted amino oxadiazole 25 can beprepared by first reacting the hydrazide with an appropriateisothiocyanate in a suitable organic solvent like DMF or THF attemperatures ranging from 25 to 100° C. The intermediate can be isolatedor can be cyclized directly with the treatment of EDCl or othercarbodiimide in suitable organic solvent like THF or DMF at temperaturesranging from room temperature to 80° C.

In Scheme 5, the preparation of keto benzimidazole derivatives isillustrated. In step 1, the methyl ester is converted to the benzylalcohol by standing reductive methods, preferably LAN in THF at 0° C. orNaBH₄ in EtOH:THF at room temperature. Oxidation to the aldehyde can beaccomplished in step 2 using MnO₂ in acetone THF at 50° C. In step 3,organometallic reagents, such as organolithium reagents and Grignardreagents, can be added to the aldehyde in THF at low temperature (e.g.,−78° C.) to give the substituted benzyl alcohol. The keto derivativescan be prepared in step 4 by oxidation of the benzyl alcohol understandard conditions such as Swern or Dess-Martin oxidation.

The compounds of the present invention may have asymmetric carbon atoms.Diastereomeric mixture can be separated into their individualdiastereomers on the basis of their physical chemical differences bymethods known to those skilled in the art, for example, bychromatography or fractional crystallization. Enantiomers can beseparated by converting the enantiomer mixture into a diastereomericmixture by reaction with an appropriate optically active compound (e.g.,alcohol), separating the diastereomers and converting (e.g.,hydrolyzing) the individual diastereomers to the corresponding pureenantiomers. All such isomers, including diastereomeric mixtures andpure enantiomers are considered as part of the invention.

The activity of the compounds of the present invention may be determinedby the following procedure. N-terminal 6 His-tagged, constitutivelyactive MEK1 (2-393) is expressed in E. coli and protein is purified byconventional methods (Ahn et al., Science 1994, 265, 966-970). Theactivity of MEK1 is assessed by measuring the incorporation ofγ-³³P-phosphate from γ-³³P-ATP onto N-terminal His tagged ERK2, which isexpressed in E. coli and is purified by conventional methods, in thepresence of MEK1. The assay is carried out in 96-well polypropyleneplane. The incubation mixture (100 μL) comprises of 25 mM Hepes, pH 7.4,10 mM MgCl₂, 5 mM β-glycerolphosphate, 100 μM Na-orthovanadate, 5 mMDTT, 5 nM MEK1, and 1 μM ERK2. Inhibitors are suspended in DMSO, and allreactions, including controls are performed at a final concentration of1% DMSO. Reactions are initiated by the addition of 10 μM ATP (with 0.5μCi γ-³³P-ATP/well) and incubated at ambient temperature for 45 minutes.Equal volume of 25% TCA is added to stop the reaction and precipitatethe proteins. Precipitated proteins are trapped onto glass fiber Bfilterplates, and excess labeled ATP washed off using a Tomtec MACH IIIharvestor. Plates are allowed to air-dry prior adding 30 μL/well ofPackard Microscint 20, and plates are counted using a Packard TopCount.In this assay, compounds of the invention exhibited an IC₅₀ of less than50 micromolar.

The following compounds were evaluated in the above assay and found tobe active. Compound # Activity  8n active 11b active 11c active 11pactive 18i active 29c active 29i active 29s active 29t active 29bbactive 29lll active 29mmm active

Administration of the compounds of the present invention (hereinafterthe “active compound(s)”) can be effected by any method that enablesdelivery of the compounds to the site of action. These methods includeoral routes, intraduodenal router, parenteral injection (includingintravenous, subcutaneous, intramuscular, intravascular or infusion),topical, and rectal administration.

The amount of the active compound administered will be depended on thesubject being treated, the severity of the disorder or condition, therate of administration, the disposition of the compound and thediscretion of the prescribing physician. However, an effective dosage isin the range of about 0.001 to about 100 mg per kg body weight per day,preferably about 1 to about 35 mg/kg/day, in single or divided doses.For a 70 kg human, this would amount to about 0.05 to 7 g/day,preferably about 0.05 to about 2.5 g/day. In some instances, dosagelevels below the lower limit of the aforesaid range may be more thanadequate, while in other cases still larger doses may be employedwithout causing any harmful side effect, provided that such larger dosesare first divided into several small doses for administration throughoutthe day.

The active compound may be applied as a sole therapy or may involve oneor more other anti-tumor substances, for example those selected from,for example, mitotic inhibitors, for example vinblastine: alkylatingagents, for example cisplatin, carboplatin and cyclophosphamide,anti-metabolites, for example 5-fluorouracil, cytosine arabinside andhydroxyurea, or, for example, one of the preferred anti-metabolitesdisclosed in European Patent Application No. 239362 such asN-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamicacid; growth factor inhibitors; cell cycle inhibitors; intercalatingantibiotics, for example adriamycin and bleomycin; enzymes, for example,interferon; and anti-hormones, for example anti-estrogens such asNolvadex™ (tamoxifen) or, for example anti-androgens such as Casodex™(4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide).Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of treatment.

The pharmaceutical composition may, for example, be in a form suitablefor oral administration as a tablet, capsule, pill, powder, sustainedrelease formulations, solution, suspension, for parenteral injection asa sterile solution, suspension or emulsion, for topical administrationas an ointment or cream or for rectal administration as a suppository.The pharmaceutical composition may be in unit dosage forms suitable forsingle administration of precise dosages. The pharmaceutical compositionwill include a conventional pharmaceutical carrier or excipient and acompound according to the invention as an active ingredient. Inaddition, it will include other medicinal or pharmaceutical agents,carriers, adjuvants, etc.

Exemplary parenteral administration forms include solutions orsuspensions of active compounds in sterile aqueous solutions, forexample, aqueous propylene glycol or dextrose solutions. Such dosageforms can be suitably buffered, if desired.

Suitable pharmaceutical carriers include inert diluents or fillers,water and various organic solvents. The pharmaceutical compositions may,if desired, contain additional ingredients such as flavorings, binders,excipients and the like. Thus for oral administration, tabletscontaining various excipients, such as citric acid may be employedtogether with various disintegrants such as starch, alginic acid andcertain complex silicates and with binding agents such as sucrose,gelatin and acacia. Additionally, lubricating agents such as magnesiumstearate, sodium lauryl sulfate and talc are often useful for tabletingpurposes. Solid compositions of a similar type may also be employed insoft and hard filled gelatin capsules. Preferred materials, therefore,include lactose or milk sugar and high molecular weight polyethyleneglycols. When aqueous suspensions or elixirs are desired for oraladministration the active compounds therein may be combined with varioussweetening or flavoring agents, coloring matters or dyes and, ifdesired, emulsifying agents or suspending agents, together with diluentssuch as water, ethanol, propylene glycol, glycerin, or combinationsthereof.

Methods of preparing various pharmaceutical compositions with a specificamount of active compound are known, or will be apparent, to thoseskilled in this art. For examples, see Remington's PharmaceuticalSciences. Mack Publishing Company, Ester, Pa., 15^(th) Edition (1975).

The examples and preparations provided below further illustrate andexemplify the compounds of the present invention and method of preparingsuch compounds. It is to be understood that the scope of the presentinvention is not limited in any way by the scope of the followingexample and preparations. In the following examples molecules with asingle chiral center, unless otherwise noted, exist as a racemicmixture. Those molecules with two or more chiral centers, unlessotherwise noted, exist as a racemic mixture of diastereomers. Singleenantiomers/diastereomers may be obtained by methods known to thoseskilled in the art.

The disclosures in this application of all articles and references,including patents, are incorporated herein by reference.

The invention is illustrated further by the following examples which arenot to be construed as limiting the invention is scope or spirit to thespecific procedures described in them.

The starting materials and various intermediates may be obtained fromcommercial sources, prepared from commercially available organiccompounds, or prepared using well known synthetic methods.

Representative examples of methods for preparing intermediates of theinvention are set forth below.

EXAMPLES Example 1

7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (11a)

Step A: 2,3,4-Trifluoro-5-nitro-benzoic acid 2

A 3 liter three neck round bottom flask is charged with 125 mL H₂SO₄.Fuming nitric acid is added (8.4 mL, 199 mmol) and the mixture gentlystirred. 2,3,4-Trifluorobenzoic acid 1 (25 g, 142 mmol) is added in 5 gportions over 90 minutes. The dark brownish yellow solution is stirredfor 60 minutes at which time the reaction is complete. The reactionmixture is poured into 1 liter of an ice:water mixture and extractedwith diethyl ether (3×600 mL). The combined organic extracts are dried(MgSO₄) and concentrated under reduced pressure to give a yellow solid.The solid is suspended in hexanes and stirred for 30 minutes after whichtime it is filtered to give 29 g (92%) of clean desired product as anoff-yellow solid: MS APCI (−) m/z 200 (M−1) detected.

Step B: 4-Amino-2,3-difluoro-5-nitro-benzoic acid 3

Ammonium hydroxide solution (˜30% in water) (35 mL, 271 mmol) is addedto a solution of 2,3,4-trifluoro-5-nitro-benzoic acid 2 (15 g, 67.8mmol) in 30 mL water at 0° C. with stirring. Upon completion of ammoniumhydroxide addition the reaction mixture is warmed at room temperaturewith stirring. After 2.5 hours, the reaction mixture is cooled to 0° C.and concentrated HCl is carefully added until pH of reaction mixture isnear 0. The reaction mixture is diluted with water (30 mL) and extractedwith diethyl ether (3×50 mL). The combined organic extracts are dried(MgSO₄) and concentrated under reduced pressure to give 14 g (95%) ofpure desired product: MS APCI (−) m/z 217 (M−1) detected.

Step C: 4-Amino-2,3-difluoro-5-nitro-benzoic acid methyl ester 4

A 2 M solution of TMS diazomethane in hexanes (6.88 mL, 13.75 mmol) isadded to a suspension of 4-amino-2,3-difluoro-5-nitro-benzoic acid 3(2.00 g, 9.17 mmol) in 25 mL of 4:1 THF:MeOH at 0° C. under nitrogenatmosphere. Upon completion of addition, reaction mixture is warmed toroom temperature. After 0.5 hours, excess TMS diazomethane is destroyedby the careful addition of acetic acid. The reaction is thenconcentrated under reduced pressure and dried in vacuo to give 1.95 g(92%) of pure desired product: MS APCI (−) m/z 231 (M−1) detected.

Step D: 4-Amino-3-fluoro-5-nitro-2-o-tolylamino-benzoic acid methylester 5a

4-Amino-2,3-difluoro-5-nitro-benzoic acid methyl ester 4 (12.0 g, 51.7mmol) is suspended in xylenes (60 mL) and ortho-toluidine is added (55.2mL, 518 mmol). The reaction mixture is heated to reflux with stirringunder a nitrogen atmosphere. After 36 hours, the reaction mixture iscooled to room temperature, diluted with diethyl ether and washed with10% aqueous HCl solution. The aqueous washing are extracted with diethylether. The combined organic extracts are concentrated under reducedpressure. The residue is dissolved in methylene chloride and filteredthrough silica gel in a fritted funnel, rising with methylene chloride.Three fractions are recovered. The first (2 liter) is nearly clean byHPLC. The second (1 liter) and third (1 liter) fraction are onlypartially pure. The first fraction is concentrated under reducedpressure and triturated with diethyl ether to give 11.2 g (68%) of cleandesired product as a bright yellow solid: MS APCI (−) m/z 318 (M−1)detected.

Step E: 7-Fluoro-6-o-tolylamino-1H-benzoimidazole-5-carboxylic acidmethyl ester 7a

4-Amino-3-fluoro-5-nitro-2-o-tolylamino-benzoic acid methyl ester 5a(1.57 g, 4.92 mmol), formic acid (25 mL, 26.5 mmol) and 20% Pd(OH)₂/C(1.57 g, 2.95 mmol) in 25 mL EtOH are heating with stirring to 95° C.After 16 hours, the reaction mixture is cooled to room temperature and0.5 g 20% Pd(OH)₂/C and 10 mL formic acid added. The reaction mixture isheated to 95° C. with stirring. After 16 hours, the reaction mixture iscooled to room temperature and filtered through Celite rinsing withEtOH. The filtrate is concentrated under reduced pressure until thedesired product precipitates. The desired product is collected byfiltration. The filtrate is concentrated again until more desiredproduct precipitates. The product is collected by filtration. RepeatedEtOH concentration, product filtration several times. recovered 1.09 g(74%) pure desired product: MS APCI (+) m/z 300 (M+1) detected; MS APCI(−) m/z 298 (M−1) detected.

Step F:7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid ester 8a

7-Fluoro-6-o-tolylamino-1H-benzoimidazole-5-carboxylic acid methyl ester7a (2.00 g, 6.68 mmol) is suspended in 1:1 THF:MeOH mixture (60 mL) andcooled to −78° C. under a nitrogen atmosphere. A solution of NBS (1.20g, 6.75 mmol) in 1:1 THF/MeOH (5 mL) is added following by MeOH (5 mL)solution of TsOH.H₂O (1.9 g, 10.0 mmol). After 30 minutes, the reactionmixture is warmed to 0° C. and then after 1 hour warmed to rt. After 16hours, more NBS (0.12 g, 0.67 mmol) is added and the reaction mixture isallowed to stir for 3 hours. The reaction mixture is quenched by theaddition of 10% Na_(S) ₂O₄ solution. After 30 minutes, the reactionmixture is diluted with water and ethyl acetate and the layersseparated. The aqueous layer is extracted with ethyl acetate. Thecombined organic extracts are dried (Na₂SO₄) and concentrated underreduced pressure. The recovered solid is triturated with methylenechloride to give 2.00 g (79%) pure desired product: MS APCI (+) m/z 380,378 (M+1 Br pattern) detected.

Step G:7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid 10a

7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid methyl ester 8a (63 mg, 0.167 mmol) is suspended in MeOH (1.5 mL)and 20% NaOH (400 μl) is added. After 16 hours, the reaction is cooledto 0° C. and 1 N NCl solution is added dropwise until pH is 2 to 3. Thereaction mixture is diluted with ethyl acetate and water and the layersseparated. The organic layer is washed with brine, dried (Na₂SO₄) andconcentrated under reduced pressure to give 58 mg (95%) of desiredproduct: MS APCI (+) m/z 366, 364 (M+1 Br pattern) detected; MS APCI (−)m/z 364, 362 (M−1 Br pattern) detected.

Step H:7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11a

7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid 10a (48 mg, 0.132 mmol) is dissolved in 1:1 THF:methylene chloride(1 mL) and Hunig's base (0.23 μl, 1.32 mmol) is added followed by PyBOP(82 mg, 0.158 mmol). After a few minutes, cyclopropyl methylhydroxylamine hydrochloride (20 mg, 0.158 mmol) (WO 0042022) is added.After the reaction is complete, the mixture is partitioned betweenmethylene chloride and saturated NaHCO₃ solution. The layers areseparated and the organic layer is washed with saturated NaHCO₃ andbrine. The organic layer is dried (Na₂SO₄) and concentrated underreduced pressure. After purification by FCC (elute with 20:1 methlenechloride:MeOH), 25 mg (45%) of pure desired product is isolated: MS ESI(+) m/z 435, 433 (M+1 Br pattern) detected; MS ESI (−) m/z 433, 431 (M−1Br pattern) detected; ¹H NMR (400 MHz, CDCl₃) δ8.15 (s, 1H), 8.02 (s,1H), 7.28 (s, 1H), 7.43 (d, 1H), 7.07 (dd, 1H), 6.36 (m, 1H), 3.70 (d,2H), 2.38 (s, 3H), 0.86 (m, 1H), 0.41 (m, 2H), 0.13 (m, 2H); ¹⁹F NMR(375 MHz, CDCl₃)−134.05 (s).

Example 2

7-Fluoro-6-phenylamino-3H-benzoimidazole-5-carboxylic acid methyl ester(27a)

Step A: 4-Amino-3-fluoro-5-nitro-2-phenylamino-benzoic acid methyl ester26a

4-Amino-2,3-difluoro-5-nitro-benzoic acid methyl ester 4 (23.48 g, 101.1mmol), the product of Example 1, Step C, is suspended in xylenes (125mL) and aniline (92 mL, 101.1 mmol) is added. The reaction mixture isstirred at 125° C. for 16 hours under N₂. The reaction mixture is cooledto room temperature and solids precipitate out of solution. The solidsare collected by filtration and are washed with xylenes and then diethylether. Recovered 22.22 g (72.78 mmol) of yellow solid which is puredesired product. The filtrate is concentrated under reduced pressure,redissolved in methylene chloride and flushed through a plug of silicagel eluting with methylene chloride. The desired fractions areconcentrated under reduced pressure to give a brown solid which istriturated with diethyl ether to give 5.47 g (17.91 mmol) of yellowsolid which is pure desired product. Combined product yield is 27.69 g(90%). MS APCI (−) m/z 304 (M−1) detected.

Step B: 7-Fluoro-6-phenylamino-3H-benzoimidazole-5-carboxylic acidmethyl ester 27a

4-Amino-3-fluoro-5-nitro-2-phenylamino-benzoic acid methyl ester 26a(16.70 g, 54.71 mmol), formic acid (250 mL, 6.63 mmol) and 20% Pd(OH)₂/C(9.00 g, 16.91 mmol) in ethanol (250 mL) are stirred at 40° C. for twohours under N₂ and then at 95° C. for 16 hours. The reaction mixture iscooled to room temperature and filtered through Celite rinsing withethyl acetate. The filtrate is concentrated under reduced pressure togive a yellow solid. The solid is triturated with diethyl ether to give13.47 g (86%) of the desired product as a tan solid. MS APCI (+) m/z 286(M+1) detected; MS APCI (−) m/z 284 (M−1) detected.

Example 3

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester (8b)

Step A: 6-(4-Bromo-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 28a

7-Fluoro-6-phenylamino-3H-benzoimidazole-5-carboxylic acid methyl ester27a (4.99 g, 17.51 mmol) is dissolved in N,N-dimethylformamide (275 mL).N-bromosuccinimide (3.15 g, 17.70 mmol) is added as a solid and thereaction mixture is stirred at room temperature under N₂. After 30minutes, the reaction mixture is quenched by the addition of aqueoussaturated sodium bisulfite solution. The reaction mixture is then pouredinto a separatory funnel, diluted with water and ethyl acetate and thelayers separated. The aqueous layer is extracted with ethyl acetate. Thecombined organic extract are washed three times with water, once withbrine and then are dried (Na₂SO₄) and concentrated under reducedpressure to yield 6.38 g (100%) of the pure desired product as a tansolid. MS ESI (+) m/z 364, 366 (M+ Br pattern) detected.

Step B:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b

6-(4-Bromo-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylic acidmethyl ester 28a (6.38 g, 17.51 mmol) is dissolved inN,N-dimethylformamide (275 mL). N-chlorosuccinimide (2.36 g, 17.70 mmol)is added as a solid and the reaction mixture is stirred at roomtemperature under N₂ until the reaction is complete (5-6 days). Thereaction mixture is quenched by the addition of aqueous saturated sodiumbisulfite solution to give a suspension. The resulting solids arecollected by filtration, washed with water and diethyl ethyl and driedunder reduced pressure to yield 6.07 g (87%), of the pure desiredproduct as a beige solid. MS ESI (+) m/z 398, 400 (M+ Br pattern)detected.

Example 4

6-(2,4-Dichloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester (8c)

7-Fluoro-6-phenylamino-3H-benzoimidazole-5-carboxylic acid methyl ester27a (1.00 g, 3.51 mmol) is suspended in 1:1 tetrahydrofuran/methanol (20mL) and cooled to −78° C. under N₂. TsOH.H₂O (3.00 g, 10.50 mmol) isadded followed by N-chlorosuccinimide (0.95 g, 7.08 mmol). After 10minutes, the reaction mixture is warmed to 0° C. to give a solution andthen 30 minutes later warmed to room temperature. After stirring for 16hours, the reaction is complete. The reaction mixture is quenched by theaddition of aqueous saturated sodium bisulfite solution and diluted withethyl acetate and water and the layers separated. The aqueous layer isextracted with ethyl acetate. The combined organic extract are washedwith brine, dried (Na₂SO₄) and concentrated under reduced pressure. Theresulting solid residue is triturated with methylene chloride to yield awhite solid which is collected by filtration to yield 1.05 g (85%) ofthe pure desired product. MS ESI (+) m/z 355, 357 (M+ Cl pattern)detected.

Example 5

6-(4-Bromo-2-fluoro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester (8d)

Step A: 4-Amino-3-fluoro-2-(2-fluoro-phenylamino)-5-nitro-benzoic acidmethyl ester 5b

4-Amino-2,3-difluoro-5-nitro-benzoic acid methyl ester 4 (1.50 g, 6.46mmol) is suspended in xylenes (7.5 mL) and 2-fluoro-phenylamine (6.24mL, 64.6 mmol) is added. The reaction mixture is stirred at 140° C.under N₂. After stirring for 6 days, the reaction is complete. Thereaction mixture is cooled to room temperature and diluted withmethylene chloride and filtered through a silica gel plug eluting withmethylene chloride (1L) to give an orange filtrate. The filtrate isconcentrated to dryness and then triturated with diethyl ether to yielda bright yellow solid. The trituration is repeated. The yellow solid iscollected to yield 1.08 g (52%) of the pure desired product. MS APCI (−)m/z 322 (M−1) detected.

Step B:6-(4-Bromo-2-fluoro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8d

4-Amino-3-fluoro-2-(2-fluoro-phenylamino)-5-nitro-benzoic acid methylester 5b is converted by the reduction/cyclization and brominationprocedures already described to yield the described product. MS ESI (+)m/z 382, 384 (M+, Br pattern) detected.

Example 6

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester (8e)

7-Fluoro-6-o-tolylamino-3H-benzoimidazole-5-carboxylic acid methyl ester7a is converted by the procedure already described for bromination,except N-chlorosuccinimide is used instead of N-bromosuccinimide, toyield the desired product. MS ESI (+) m/z 334, 336 (M+, Cl pattern)detected.

Example 7

7-Fluoro-6-(2-methyl-4-trifluoromethoxy-phenylamino)-3H-benzoimidazole-5-carboxylicacid methyl ester (8f)

Step A.4-Amino-3-fluoro-2-(2-methyl-4-trifluoromethoxy-phenylamino)-5-nitro-benzoicacid methyl ester 12a

4-Amino-2,3-difluoro-5-nitro-benzoic acid methyl ester 4 (0.50 g, 2.15mmol) is suspended in xylenes (3 mL) and2-methyl-4-trifluoromethoxy-phenylamine (1.00 g, 5.23 mmol) is added.The reaction mixture is stirred at 140° C. under N₂. After stirring for7 days, the reaction is a mixture of starting material and product. Thereaction mixture is cooled to room temperature. The reaction mixture ispoured into a separatory funnel and diethyl ether and 10% aqueous HClare added and the layers separated. The aqueous phase is extracted withthree portions of diethyl ether. The combined diethyl ether layers aredried (MgSO₄) and concentrated under reduced pressure. The residue isredissolved in methylene chloride and flushed through a plug of silicagel eluting with methylene chloride. The filtrate is concentrated underreduced pressure to give a bright yellow solid. The solid is washed withdiethyl ether and the filtrate is concentrated under reduced pressureand the residue is further purified by FCC (eluting with 100% methylenechloride) to yield 0.39 g (45%) of the desired pure product as a yellowsolid. MS APCI (−) m/z 402 (M−1) detected.

Step B.7-Fluoro-6-(2-methyl-4-trifluoromethoxy-phenylamino)-3H-benzoimidazole-5-carboxylicacid methyl ester 8f

4-Amino-3-fluoro-2-(2-methyl-4-trifluoromethoxy-phenylamino)-5-nitro-benzoicacid methyl ester 12a is converted by the reduction/cyclizationprocedure already described to yield the desired product. MS APCI (+)m/z 384 (M+1) detected; MS APCI (−) m/z 382 (M−1) detected.

Example 8 Preparation of Hydroxylamines

Hydroxylamines useful for synthesizing compounds of the presentinvention mat be prepared as follows.

(i) O-(2-Methoxy-ethyl)-hydroxylamine

Step A: 2-(2-Methoxy-ethoxy)-isoindole-1,3-dione

DEAD (10 mL, 63 mmol) is added to a mixture of 2-methoxyethanol (5.0 mL,63 mmol), PPh₃ (17 g, 63 mmol), and N-hydroxyphthalimide (10 g, 62 mmol)in THF (170 mL). The resulting orange solution is stirred 16 hours atroom temperature. The reaction mixture is concentrated in vacuo, and thesolids are filtered washing with CHCl₃. The filtrate is concentratedagain, and the solids are filtered washing with CHCl₃. This process isrepeated until no precipitate forms. The final yellowish solids arerecrystallized from EtOH to give the desired product (7.7 g, 55%).

Step B: O-(2-Methoxy-ethyl)-hydroxylamine

To a solution of 2-(2-methoxy-ethoxy)-isoindole-1,3-dione (7.7 g, 35mmol) in CH₂Cl₂ (30 mL) at room temperature is added methylhydrazine(2.0 mL, 36 mmol). The resulting solution is stirred for 16 hours atroom temperature. The white solids are filtered off. The solvent iscarefully distilled off under reduced pressure, then the concentrate isdistilled under vacuum (20 torr, 57-58° C.) to afford the desiredproduct (2.2 g, 68%).

(ii) The following hydroxylamines are prepared as described above usingthe appropriate alcohols. The isoindole-1,3-dione intermediates arepurified by flash chromatography.

O-(2-Isobutoxy-ethyl)-hydroxylamine is used directly withoutpurification.

O-(2-Pyrrolidin-1-yl-ethyl)-hydroxylamine is used directly withoutpurification.

O-(2-Piperidin-1-yl-ethyl)-hydroxylamine is purified by Kugelrohrdistillation (chamber temperature 140° C., 1 torr).

O-(2-Methylsulfanyl-ethyl)-hydroxylamine is purified by vacuumdistillation (76-78° C., 20 torr).

O-(2-Phenylsulfanyl-ethyl)-hydroxylamine is used directly withoutpurification.

O-(3-Methylsulfanyl-propyl)-hydroxylamine is used directly withoutpurification.

(iii) The following hydroxylamines are prepared from the appropriateisoindole-1,3-dione by oxidation using oxone (Tetrahedron Lett. 1981,22, 1287), and then deprotection as described above.

O-(2-Methanesulfonyl-ethyl)-hydroxylamine is used directly withoutpurification.

O-(2-Benzenesulfonyl-ethyl)-hydroxylamine is purified by flashchromatography (1% MeOH in CH₂Cl₂).

O-(3-Methanesulfonyl-propyl)-hydroxylamine is used directly withoutpurification.

O-(3-Phenylsulfanyl-propyl)-hydroxylamine is prepared fromPhSCH₂CH₂CH₂Br and N-hydroxyphthalimide by the present procedure WO0018790 and then is deprotected by the procedure described above andused directly without purification.

(iv)

O-(3-Benzenesulfonyl-propyl)-hydroxylamine is prepared from the aboveisoindole-1,3-dione through its oxidation with oxone followed bydeprotection as described above and is purified by flash chromatography(100% CH₂Cl₂ to 2% MeOH in CH₂Cl₂).

(v) O-(2-Morpholin-4-yl-ethyl)-hydroxylamine dihydrochloride

Step A: O-(2-Bromo-ethyl)-hydroxylamine hydrobromide

2-(2-Bromo-ethoxy)-isoindole-1,3-dione is prepared from1,2-dibromoethane and N-hydroxyphtalimide as described in WO 0018790,and is then subjected to the procedure in J. Org. Chem. 1963, 28, 1604to yield the desired product.

Step B: (2-Bromo-ethoxy)-carbamic acid tert-butyl ester

To a solution of O-(2-bromo-ethyl)-hydroxylamine hydrobromide (100 mg,0.45 mmol) and di-t-butyl dicarbonate (110 mg, 0.49 mmol) in CH₂Cl₂ (1mL) at room temperature is added Et₃N (0.08 mL, 0.56 mmol). Theresulting suspension is stirred for 16 hours room temperature. Thereaction mixture is diluted with EtOAc, washed with 1 N aq HCl andbrine, dried over MgSO₄, filtered, concentrated, and purified by flashchromatography (100% CH₂Cl₂) to give the desired product (81 mg, 75%).

Step C: (2-Morpholin-4-yl-ethoxy)-carbamic acid tert-butyl ester

To a solution of (2-bromo-ethoxy)-carbamic acid tert-butyl ester (232mg, 1.05 mmol) in DMF (2 mL) at room temperature is added morpholine(0.14 mL, 1.6 mmol). The reaction mixture is stirred for 7 h at 50° C.The reaction mixture is diluted with EtOAc, and washed with water. Theorganic layer is dried over MgSO₄, filtered, concentrated, and purifiedby flash chromatography (2% MeOH in CH₂Cl₂) to give the desired product(118 mg, 46%): MS APCI (+) m/z 247 detected.

Step D: O-(2-Morpholin-4-yl-ethyl)-hydroxylamine dihydrochloride

To a solution of (2-morpholin-4-yl-ethoxy)-carbamic acid tert-butylester (118 mg, 0.48 mmol) in MeOH (1 mL) is added 4 M dioxane solutionof HCl (2.4 mL, 9.60 mmol) at room temperature. The resulting solutionis stirred for 16 hours at room temperature. After addition ofadditional HCl (2.4 mL) followed by stirring for 4 hours, the reactionmixture is concentrated in vacuo to give yellow solids (82 mg, 78%).

The isoindole-1,3-dione intermediates of the following hydroxylaminesare prepared from the appropriate alkyl halide and N-hydroxyphthalimideby the procedure described within J. Hetercyclic Chem. 2000, 37,827-830. The isoindole-1,3-diones are deprotected by the proceduredescribed above: O-but-3-enyl-hydroxylamine;O-(tetrahydro-furan-2-ylmethyl)-hydroxylamine;O-(3-methoxy-propyl)-hydroxylamine; andO-(3-benzyloxy-propyl)-hydroxylamine.

(vii) The following hydroxylamines are prepared as described in WO02/06213; O-(2-vinyloxy-ethyl)-hydroxylamine;2-aminooxy-2-methyl-propan-1-ol; 1-aminooxy-2-methyl-propan-2-ol;3-aminooxy-propan-1-ol; and (2-aminooxy-ethyl)-methyl-carbamic acidtert-butyl ester.

Example 9

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (11b)

Step A: 4-Amino-2-(2-chloro-phenylamino)-3-fluoro-5-nitro-benzoic acidmethyl ester 5b.

4-Amino-2,3-difluoro-5-nitro-benzoic acid methyl ester 4 (2.00 g, 8.62mmol) is suspended in xylenes (15 mL) and 2-chloro aniline (9.06 mL,86.15 mmol) is added. The reaction mixture is heated to 140° C. under anitrogen atmosphere. After 6 days, the reaction mixture is cooled toroom temperature, and diluted with ethyl acetate. The reaction mixtureis washed with water, 10% HCl solution and brine. The organic layer isdried (MgSO₄) and concentrated under reduced pressure. The crude productis triturated with diethyl ether, twice, to give 0.35 g (12%) puredesired product as a brownish solid.

Step B: 4,5-Diamino-2-(2-chloro-phenylamino)-3-fluoro-benzoic acidmethyl ester 6a.

4-Amino-2-(2-chloro-phenylamino)-3-fluoro-5-nitro-benzoic acid methylester 5b (0.30 g, 0.88 mmol) is suspended in AcOH (5 mL) and zinc dust(0.29 g, 4.42 mmol) is added. After 15 minutes, the reaction iscomplete. The reaction mixture is diluted with ethyl acetate andfiltered through Celite. The filtrate is washed with water, saturatedNaHCO₃, 10% K₂CO₃ and brine. The organic layer is dried (MgSO₄) andconcentrated under reduced pressure to give 0.13 g (48%) pure desiredproduct as an whitish brown foam.

Step C: 6-(2-Chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 7b.

4,5-Diamino-2-(2-chloro-phenylamino)-3-fluoro-benzoic acid methyl ester6a (0.125 g, 0.404 mmol) is suspended in EtOH (2 mL) and formamidineacetate (63 mg, 0.605 mmol) is added. The reaction mixture is heated toreflex. After 16 hours, the reaction mixture is cooled to rt and dilutedwith ethyl acetate. The organic layer is washed with water, saturatedNaHCO₃, 10% K₂CO₃ and brine. The organic layer is dried (MgSO₄) andconcentrated under reduced pressure to give 0.109 g (85%) pure desiredproduct.

Step D:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b.

6-(2-Chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylic acidmethyl ester 7b (55 mg, 0.172 mmol) is dissolved in 1:1 THF:MeOH (2 mL)and cooled to −78° C. under an atmosphere of nitrogen. TsOH.H₂O (49 mg,0.258 mmol) is added followed by NBS (31 mg, 0.174 mmol). After 10minutes, the reaction mixture is warmed to 0° C. and then 2 hours laterwarmed to rt. After 16 hours, the reaction mixture is quenched by theaddition of 10% Na₂S₂O₃ and diluted with ethyl acetate and water. Thelayers are separated and the aqueous layer is extracted with ethylacetate. The combined organic extract are dried (MgSO₄) and concentratedunder reduced pressure. The product is triturated with methylenechloride to dive 58 mg (85%) of pure desired product as a tan solid.

Step E:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid 10b.

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b (58 mg, 0.146 mmol) is suspended in EtOH (2 mL) and1 mL 2 N NaOH is added. After 16 hours, the reaction mixture is dilutedwith ethyl acetate, water, and 10% HCl solution. The layers areseparated and the organic layer is washed with brine. The organic layeris dried (MgSO₄) and concentrated under reduced pressure. Triturationwith MeOH gives 22 mg (39%) pure desired product.

Step F:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (11b).

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid 10b (22 mg, 0.057 mmol) is dissolved in DMF (1 mL) and HOBt (9 mg,0.063 mmol) followed by triethyl amine (18 μl, 0.132 mmol) is added.Cyclopropyl methyl hydroxylamine hydrochloride (8 mg, 0.062 mmol) isadded following by EDCl (14 mg, 0.074 mmol). After 16 hours, thereaction mixture is diluted with ethyl acetate and water and the layersseparated. The organic layer is washed with saturated NH₄Cl, brine,saturated NaHCO₃, water and brine. The organic layer is dried (MgSO₄)and concentrated under reduced pressure to give 23 mg (80%) pure desiredproduct. MS APCI (+) m/z 455, 453 (M+ Br pattern) detected; MS APCI (−)m/z 453, 451 (M− Br pattern) detected; ¹H NMR (400 MHz, DMSO-d₆) δ 11.69(broad s, 1 H), 8.43 (s, 1H), 7.62 (d, 1H), 7.28 (dd, 1H), 6.42 (m, 1H),3.63 (d, 2H), 1.03 (m, 1H), 0.48 (m, 2H), 0.19 (m, 2H); ¹⁹F NMR (376MHz, DMSO−d₆)−132.95 (s).

The following compounds are prepared by methods similar to thosedescribed in Example 1 and in this Example 9 by using the appropriatecarboxylic acid and the appropriate hydroxylamine: 8g

8h

8i

8j

8k

8l

8m

8n

Example 10

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide (29c)

Step A.6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid methyl ester 9a and6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-1-methyl-1H-benzoimidazole-5-carboxylicacid methyl ester.

A solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b (150 mg, 0.38 mmol), iodomethane (28 μL, 0.45 mmol)and potassium carbonate (78 mg, 0.56 mmol) in dimethylformamide (1.5 mL)is stirred at 75° C. for one hour. The reaction mixture is diluted withethyl acetate, washed with saturated aqueous potassium carbonate (2×),brine, and dried (Na₂SO₄). Flash column chromatography (20:1 methylenechloride/ethyl acetate) provides 56 mg (36%) of the more mobile6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid methyl ester 9a as a white solid. ¹⁹F NMR (376 MHz, CD₃OD)−133.5(s). MS APCI (+) m/z 412, 414 (M+, Br pettern) detected. Also isolatedin 54 mg (35%) of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-1-methyl-1H-benzoimidazole-5-carboxylicacid methyl ester as a white solid. ¹⁹ F NMR (376 MHz, CD₃OD)−139.9 (s).MS APCI (+) m/z 412, 414 (M+, Br pattern) detected.

Step B.6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid 10c

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid methyl ester 9a (56 mg, 0.14 mmol) is dissolved into 2:1 THF/water(3 mL) and NaOH (0.55 mL, 1.0 M aqueous solution, 0.55 mmol) is added.After stirring for two hours the reaction is reduced to one quarterinitial volume via rotary evaporation and the remainder diluted to 50 mLwith water. The aqueous solution is acidified to pH 2 by the addition of1.0 M aqueous HCl and extracted with 1:1 tetrahydrofuran/ethyl acetate(3×),dried (Na₂SO₄) and concentrated under reduced pressure to provide43 mg (79%) pure carboxylic acid as an off white solid. MS ESI (+) m/z397, 398 (M+, Br pattern) detected.

Step C:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-vinyloxy-ethoxy)-amide 29a

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid 10c (2.00 g, 5.0 mmol), O-(2-vinyloxy-ethyl)-hydroxylamine (0.776g, 7.5 mmol), HOBt (0.88 g, 6.5 mmol), triethylamine (1.61 mL, 2.3 mmol)and EDCl (1.3 g, 6.5 mmol) are dissolved in dimethylformamide (52 mL)and stirred at room temperature for 48 hours. The reaction mixture isdiluted with ethyl acetate, washed with water (3×), saturated potassiumcarbonate (2×), saturated ammonium chloride (2×), brine, dried (Na₂SO₄)and concentrated under reduced pressure to an off-white solid.Trituration of the solid with diethyl ether provides 2.18 g (90%)desired product as an off-white solid. MS ESI (+) m/z 483, 485 (M+ Brpattern) detected.

Step D:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide 29c

Hydrochloric acid (14 mL, 1.0 M aqueous solution, 14 mmol) is added to asuspension of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-vinyloxy-ethoxy)-amide 29a (2.18 g, 4.50 mmol) in ethanol (50mL) and the reaction mixture allowed to stir for 24 hours. The reactionmixture is concentrated to dryness by rotary evaporation and the solidspartitioned between 3:1 ethyl acetate/tetrahydrofuran and saturatedpotassium carbonate. The aqueous phase is extracted with 3:1 ethylacetate/tetrahydrofuran (3×), the combined organics dried (Na₂SO₄), andconcentrated to provide 2.11 g (100%)6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide as an off-white solid. MS ESI (+)m/z 457,459 (M+, Br pattern) detected. ¹H NMR (500 MHz, MeOH-d₄) δ 8.26 (s, 1H),7.78 (s, 1H), 7.57 (d, 1H), 7.24 (dd, 1H), 6.40 (dd, 1H), 3.86 (s, 3H),3.79 (m, 2H), 3.49 (m, 2H). ¹⁹F NMR (376 MHz, MeOH-d₄)−133.68 (s).

Example 11

The following compounds are prepared by methods similar to thosedescribed in Example 10 by using methyl ester 8b and the appropriatealkylating agent (Step A) and the appropriate hydroxylamine (Step C):29d

29e

29f

29g

29h

29i

29j

29k

29l

29m

29n

29o

29p

29q

29r

29s

29t

29u

29v

29w

29x

29y

29z

29aa

29bb

29cc

29dd

29ee

29ff

29gg

29hh

29ii

29jj

29kk

29ll

29mm

29nn

29oo

29pp

29qq

29rr

29ss

29tt

29uu

29vv

29ww

29xx

29yy

29zz

29aaa

29bbb

29ccc

29ddd

29eee

29fff

29ggg

Example 12

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2,3-dihydroxy-propoxy)-amide (29hhh)

To a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid allyloxy-amide 29tt (20 mg, 0.04 mmol) in 0.50 mL 4:1tetrahydrofuran/water is added OsO₄ (41 μL, 0.054 M solution in t-BuOH,0.002 mmol) following by NMO (7 mg, 0.06 mmol). The solution is stirredat room temperature for eight hours after which time HPLC analysisshowed complete conversion to product. The solution is then stirred withsaturated NaHSO₃ and diluted with ethyl acetate. The organic phase isdried (Na₂SO₄). Purification by FCC (DCM→20:1 DCM/MeOH) provided 16 mgdesired product as an off-white solid. Ms ESI (+) m/z 487, 489 (M+, Brpattern) detected.

Example 13

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (3,4-dihydroxy-butoxy)-amide (29iii)

6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid but-3-enyloxy-amide 29uu is subject to the dihydroxylation methoddescribed in Example 12, MS APCI (+) m/z 501, 503 (M+ Br pattern)detected.

Example 14

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-methylamino-ethoxy)-amide TFA salt (29jjj)

Prepared from(2-{[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethyl)-methyl-carbamicacid tert-butyl ester 29ww by trifluoroacetic acid deprotection inmethylene chloride. MS APCI (+) m/z 470, 472 (M+, Br pattern) detected;¹H NMR (400 MHz, CD₃OD) δ 8.31 (s, 1H), 7.74 (s, 1H), 7.51 (d, 1H), 7.19(dd, 1H), 6.39 (dd, 1H), 4.11 (m, 1H), 4.11 (m, 2H), 3,97 (s, 3H), 3.12(m, 2H), 2.72 (s, 3H); ¹⁹F NMR (376 MHz, CD₃OD)-77.41 (s, 3F), −134.79(s, 1F).

Example 15

The following compounds are prepared by methods similar to thosedescribed in Example 10 by using methyl ester 8a and the appropriatealkylating agent (Step A) and the appropriate hydroxylamine (Step C):11c

11d

11e

11f

Example 16

The following compounds are prepared by methods similar to thosedescribed in Example 10 by using methyl ester 8e and the appropriatealkylating agent (Step A) and the appropriate hydroxylamine (Step C):11g

11h

11i

11j

11k

11l

Example 17

The following compounds are prepared by methods similar to thosedescribed in Example 10 by using methyl ester 8c and the appropriatealkylating agent (Step A) and the appropriate hydroxylamine (Step C):11m

29kkk

Example 18

The following compounds are prepared by methods similar to thosedescribed in Example 10 by using methyl ester 8d and the appropriatealkylating agent (Step A) and the appropriate hydroxylamine (Step C):29lll

29mmm

Example 19

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-[4-(4-methyl-piperazin-1-yl)-butyl]-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (11o)

Step A:6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-pent-4-enyl-3H-benzoimidazole-5-carboxylicacid methyl ester 9b

7-Fluoro6-(4-bromo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid methyl ester 8a (0.915 g, 2.419 mmol) is suspended in DMF (18 mL)under an atmosphere of nitrogen. Bromopentene (0.430 mL, 3.629 mmol) andK₂CO₃ (0.502 g, 3.629 mmol) are added and the reaction mixture waswarmed to 80° C. After 1 hour, the reaction mixture is cooled to roomtemperature and poured into 100 mL of 1:1 ethyl acetate:diethyl ester.The organic layer is washed with water and brine, dried (Na₂SO₄) andconcentrated under reduced pressure. The N3 and N1 alkylated product areseparated by flash column chromatography, eluted with 20:1 methylenechloride:ethyl acetate. Complete separation of the isomers is obtainedby performing two chromatographic separations. The higher R_(f) productis the N3 product 9b, while the lower R_(f) product is the N1 product.The recovery of the N3 product 9b is 0.415 g (38%): LC/MS ESI (+) m/z448, 446 (M+1 Br pattern) detected. The recovery of the N1 product was0.486 g (45%): LC/MS ESI (+) m/z 448, 446 (M+1 Br pattern) detected.

Step B:6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-pent-4-enyl-3H-benzoimidazole-5-carboxylicacid 10d

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-pent-4-enyl-3H-benzoimidazole-5-carboxylicacid methyl ester 9b is dissolved in 1:1 THF:MeOH (10 mL) and 1 N NaOHsolution (2.3 mL) is added. After 5 h, the organic solvents are removedunder reduced pressure and the residue diluted with water and 100 mL 1:1THF:ethyl acetate. The layers are separated and the aqueous layerextracted with ethyl acetate. The combined organic extracts are dried(Na₂SO₄) and concentrated under reduced pressure to afford 0.39 g (100%)clean desired product as a light yellow solid.

Step C:6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-pent-4-enyl-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11f

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-pent-4-enyl-3H-benzoimidazole-5-carboxylicacid 10d (0/390 g, 0.902 mmol) is dissolved in 1:1 THF:methylenechloride (6 mL) and Hunig's base (0.346 mL, 1.985 mmol) is addedfollowed by PyBOP (0.563 g, 1.083 mmol). After 10 minutes, cyclopropylmethyl hydroxylamine hydrochloride (0.134 g, 1.083 mmol) is added. After16 hours, the reaction mixture is diluted with ethyl acetate and washedwith 0.1 HCl, saturated NaHCO₃, and brine. The organic layer is dried(Na₂SO₄) and concentrated under reduced pressure. The crude yellowresidue is purified by FCC eluted with ethyl acetate to give 0.315 g(70%) pure desired product as a yellow solid: MS APCI (+) m/z 503, 501(M+1 Br pattern) detected.

Step D:6-(4-Bromo-2-methyl-phenylamino)-3-(4,5-dihydroxy-pentyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylamethoxy-amide 11m

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-pent-4-enyl-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11f (0.307 g, 0.612 mmol) is dissolved in4:1 THF:water (8 mL) and 1.134 mL (0.061 mmol) of an 0.054 M OsO₄solution in t-BuOH iias added followed by NMO (0.093 g, 0.796 mmol).After 5 h, the reaction mixture is quenched by the addition of 10%NaHS₂O₃ solution. After 10 minutes, the reaction mixture is filteredthrough Celite rinsing with ethyl acetate methylene chloride. Thefiltrate is diluted with ethyl acetate and washed with 0.01 N HCl, andbrine. The organic layer is dried (Na₂SO₄) and concentrated underreduced pressure. The crude product is purified by FCC eluted with 9:1ethyl acetate:MeOH to give 0.244 g (74%) pure desired product.

Step E:6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-(4-oxo-butyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11n

To a mixture of6-(4-Bromo-2-methyl-phenylamino)-3-(4,5-dihydroxy-pentyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11m (0.244 g, 0.456 mmol), THF (5 mL) andpH 7 phosphate buffer (3 mL) is added sodium periodate (0.195 g, 0.911mmol). After 16 hours, the reaction mixture is diluted with ethylacetate and washed with NaHCO₃, and brine. The organic layers is dried(Na₂SO₄) and concentrated under reduced pressure to give an orangesolid. Purification by FCC eluted with 4:1 methylene chloride:MeOHyields 0.189 g (82%) pure desired product as a yellow solid: MS APCI (+)m/z 505, 503 (M+1 Br pattern) detected: MS APCI (−) m/z 503, 501 (M−1 Brpattern) detected.

Step F:6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-[4-(4-methyl-piperazin-1-yl)-butyl]-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11o

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3-(4-oxo-butyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11n (15 mg, 0.030 mmol) is dissolved inMeCN (500 μl) and methylpiperazine (10 μL, 0.089 mmol) is added followedby AcOH (5 μL, 0.089 mmol). After 5 minutes, tetramethylammoniumtriacetoxyborohydride (12 mg, 0.045 mmol) is added. After 5 minutes, thereaction mixture is diluted with ethyl acetate and washed with NaHCO₃and brine. The organic layer is dried (Na₂SO₄) and concentrated underreduced pressure to give 12 mg (69%) of pure desired product as a whitesolid. MS APCI (−) m/z 587, 585 (M−1 Br pattern) detected: ¹H NMR (400MHz, CDCl₃) δ 7.99 (s, 1H), 7.98 (s, 1H), 7.30 (d, 1H), 7.08 (dd, 1H),6.30 (d, 1H), 6.1 (broad singlet, 1H), 4.26 (t, 2H), 3.64 (d, 2H), 3.37(s, 1H), 2.45 (broad, 8H), 2.41 (s, 3H), 2.38 (t, 2H), 2.28 (s, 3H),1.95 (quin, 2H), 1.55 (quin, 2H), 0.98 (m, 1H), 0.50 (qt, 2H), 0.22 (qt,2H).

Example 20

The following compounds are prepared by methods similar to thosedescribed in Example 19 by using the appropriate alkenyl substitutedbenzimidazole and the appropriate amine in the reduced amination (stepF): 18a

18b

18c

18d

18e

18f

18g

18h

18i

18j

18k

18l

18m

18n

18o

18p

18q

18r

18s

18t

18u

18v

18w

18x

18y

18z

18aa

18bb

Example 21

6-(4-Bromo-2-methyl-phenylamino)-3-[4-(1,1-dioxo-1λ⁶-thiomorpholin-4-yl)-butyl]-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (18cc)

To a solution of6-(4-bromo-2-methyl-phenylamino)-7-fluoro-3-(4-thiomorpholin-4-yl-butyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 18l (8 mg, 0.014 mmol) in 1:1:1water/acetone/MeOH (1 mL) is added NMO (1.6 mg, 0.014 mmol) and osmiumtetroxide (250 μL, 0.054 M solution in t-BuOH, 0.014 mmol). Afterstirring for 24 hours, the solution is diluted with saturated sodiumthiosulfate stirred for 1 minutes and diluted with ethyl acetate. Thesolution is washed with brine (2×), dried (Na₂SO₄) and concentratedunder reduced pressure to a grey solid. FCC (10:1dichloromethane/methanol) provides 6 mg (71%) desired product as anoff-white solid. MS ESI (+) m/z 622, 624 (M+, Br pattern) detected.

Example 22

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-[4-(4-methyl-piperazin-1-yl)-butyl]-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (18dd)

A solution of6-(4-Bromo-2-chloro-phenylamino)-3-(4-chloro-butyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 18ee (10 mg, 0.018 mmol), sodium iodide(14 mg, 0.092 mmol), and 1-methyl-piperazine (10 ρL, 0.092 mmol) arestirred at 85° C. for three hours. The reaction mixture is diluted withethyl acetate and washed three times with water, washed twice withsaturated aqueous potassium carbonate, dried (Na₂SO₄) and concentratedunder reduced pressure to a yellow oil. Flash column chromatography (1:1dichloromethane/methanol followed by methanol followed by 20:1methanol/triethylamine) yields clean product (8 mg, 72%) as an off-whitefoam. MS ESI (+) m/z 607, 609 (M+, Br pattern) detected. ¹H NMR (400MHz, DMSO-d₆) δ 8.37 (s, 1H), 7.71 (s, 1H), 7.49 (d, 1H), 7.18 (dd, 1H),6.40 (dd, 1H), 4.38 (t, 2H), 3.62 (d, 2H), 2.45 (broad, 8H), 2.41 (t,2H), 2.28 (s, 3H), 1.96 (m, 2H), 1.54 (m, 2H), 1.07 (m, 1H), 0.50 (d,2H), 0.22 (d, 2H).

Example 23

The following compounds are prepared by methods similar to thosedescribed in Example 22, using an appropriate amine and primary alkylchloride. 18ff

18gg

18hh

18ii

18jj

18kk

18ll

18mm

18nn

18oo

18pp

18qq

18rr

18ss

18tt

18uu

18vv

18ww

18xx

18yy

18zz

18aaa

18bbb

18ccc

18ddd

18eee

18fff

Example 24

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3oxazol-5-ylmethyl-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (18ggg)

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3-(2-oxo-ethyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (0.020 g, 0.046 mmol) is dissolved inmethanol (2 mL). Potassium carbonate (0.013 g, 0.093 mmol) and1-isocyanomethanesulfonyl-4-methyl-benzene (0.010 g, 0.051 mmol) areadded. The reaction mixture is stirred at reflux for 16 hours under N₂,then concentrated under reduced pressure. The residue is dissolved inethyl acetate and poured into a separatory funnel and washed with waterand brine. The combined aqueous layers reextracted with ethyl acetate(2×). The combined ethyl acetate layer are dried (Na₂SO₄), andconcentrated under reduced pressure. The resulting solid was purified byflash column chromatograhy (eluting with 15:1 methylenechloride:methanol) to yield 0.011 g (50%) of the desired product. MSAPCI (+) m/z 470, 472 (M+, Cl pattern) detected; ¹H NMR (400 MHz, CDCl₃)δ 10.51 (br s, 1H), 8.07 (s, 1H), 8.02 (s, 1H), 7.89 (s, 1H), 7.23 (s,1H), 7.15 (d, 1H), 6.92 (dd, 1H), 6.31 (d, 1H), 6.11 (br, 1H), 5.45 (s,2H), 3.62 (d, 2H), 2.40 (s, 3H), 0.87 (m, 1H), 0.49 (m, 2H), 0.20 (m,2H). ¹⁹F NMR (376 MHz, CDCl₃)−134.54 (s)

Example 25

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(3-oxo-3-pyrrolidin-1-yl-propyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (18hhh)

Step A:6-(4-Bromo-2-chloro-phenylamino)-3-(2-tert-butoxycarbonyl-ethyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b (0.50 g, 1.25 mmol) is dissolved in DMF (8 mL)under N₂ and K₂CO₃ (0.26 g, 1.88 mmol) is added followed by t-butylacrylate (1.84 mL, 12.54 mmol). The reaction mixture heated to 90° C.with stirring. After 4 hours, the reaction mixture is cooled to rt anddiluted with ethyl acetate. The organic layer is washed with water (3×)and brine, dried (MgSO₄) and concentrated under reduced pressure.Purification by flash column chromotography eluted with 19:1 methylenechloride:ethyl acetate gives 0.41 g (62%) desired product.

Step B:6-(4-Bromo-2-chloro-phenylamino)-3-(2-carboxy-ethyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester TFA salt

6-(4-Bromo-2-chloro-phenylamino)-3-(2-tert-butoxycarbonyl-ethyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester (0.050 g, 0.095 mmol) is dissolved in methylenechloride (0.5 mL) and TFA (0.5 mL) is added. After 45 minutes, thereaction mixture is concentrated to dryness to give 0.49 g (88%) desiredproduct: LC/MS ESI (+) m/z 472, 470 (M+ Br pattern) detected: ¹H NMR(400 MHz, DMSO-d₆)

8.51 (s, 1H), 8.20 (s, 1H), 8.13 (s, 1H), 7.64 (d, 1H), 7.29 (dd, 1H),6.45 (dd, 1H), 4.55 (t, 2H), 2.89 (t, 2H).

Step C:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(3-oxo-3-pyrrolidin-1-yl-propyl)-3H-benzoimidazole-5-carboxylicacid methyl ester

To a solution of6-(4-bromo-2-chloro-phenylamino)-3-(2-carboxy-ethyl)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester (60 mg, 0.13 mmol) in DMF (1.8 mL) is added HOBt-H₂O(24 mg, 0.16 mmol), Et₃N (0.043 mL, 0.31 mmol), pyrrolidine (0.011 mL,0.13 mmol), and EDCl (34 mg, 0.18 mmol) at rt. The resulting yellowsolution is stirred 16 hours at rt. The reaction mixture is diluted withEtOAc and water, washed with sat'd aq NH₄Cl, brine, sat'd aq NaHCO₃, andbrine. The organic layer is dried over MgSO₄, filtered, and concentratedin vacuo to give a crude material which is purified by flashchromatography (3% MeOH in CH₂Cl₂) to afford 45 mg (67%) of the desiredproduct: MS APCI (+) m/z 523, 525 (M+, Br pattern) detected.

Step D:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(3-oxo-3-pyrrolidin-1-yl-propyl)-3H-benzoimidazole-5-carboxylicacid

To a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-(3-oxo-3-pyrrolidin-1-yl-propyl)-3H-benzoimidazole-5-carboxylicacid methyl ester (41 mg, 0.079 mmol) in THF/H₂O (1.5 mL/0.75 mL) isadded 0.20 mL (0.20 mmol) of 1 N aq LiOH at rt. The resulting solutionis stirred 16 hours. The reaction mixture is acidified with 1 N aq HCl(pH ˜2 to 3) and diluted with EtOAc. The organic layer is dried overMgSO₄, filtered, and concentrated in vacuo to give a crude product (42mg) which is directly used without further purification.

Step E:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(3-oxo-3-pyrrolidin-1-yl-propyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 18hhh

The title compound is prepared from6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-(3-oxo-3-pyrrolidin-1-yl-propyl)-3H-benzoimidazole-5-carboxylicacid and O-cyclopropylmethyl-hydroxylamine hydrochloride by the standardcoupling procedure described in Step A: MS APCI (+) m/z 578, 580 (M+, Brpattern) detected; ¹H NMR (400 MHz, DMSO-d₆) δ 11.66 (s, 1H), 8.42 (s,1H), 8.01 (s, 1H), 7.76 (s, 1H), 7.62 (s, 1H), 7.28 (d, 1H), 6.39 (890m, 1H), 4.52 (t, 2H), 3.66 (d, 2H), 3.33 (t, 2H), 3.28 (t, 2H), 2.89 (t,2H), 1.83 (m, 2H), 1.76 (m, 2H), 1.06 (m, 1H), 0.49 (m, 1H), 0.22 (m,2H); ¹⁹F NMR (376 MHz, DMSO-d₆)−132.94 (s, 1F).

Example 26

The following compounds are prepared by methods similar to thosedescribed in Example 25 using methyl ester 8b and the appropriateamines: 18iii

18jjj

18kkk

Example 27

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-hydro-ethoxy)-amide (11p)

Step A:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid methyl ester 11q

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b (0.25 g, 0.63 mmol) is dissolved inN,N-dimethylformamide (5 mL). 2-Bromomethyl-tetrahydro-pyran (0.34 g,1.88 mmol) and potassium carbonate (0.26 g, 1.88 mmol) are added and thereaction mixture is stirred at 60° C. for 12 hours under N₂. Thereaction mixture is poured into a separatory funnel, diluted with ethylacetate and water and the layers separated. The ethyl acetate is washedwith water and brine, dried (Na₂SO₄) and concentrated under reducedpressure. The resulting solid residue is triturated with diethyl etherto yield a pale yellow solid (N3 regioisomers by NMR) and a yellowfiltrate (mixture of N1 and N3 regioisomers by NMR). The solids arecollected and washed with diethyl ether to yield 0.12 g (37%) of thepure desired N3 regioisomeric product as a pale yellow solid. MS ESI (+)m/z 496, 498 (M+, Br pattern) detected.

Step B:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid 11r

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid methyl ester 11q is suspended in 4:1 tetrahydrofuran/water (2.5 mL)and aqueous 1 M LiOH is added (2.5 mL). After stirring at roomtemperature for 16 hours, the reaction mixture is homogeneous and thereaction is complete. The reaction mixture is cooled to 0° C., dilutedwith water and aqueous 2 M HCl is added dropwise until the pH of thesolution is 1-2, at which time it turns to a suspension. The reactionmixture is poured into a separatory funnel and diluted with ethylacetate/tetrahydrofuran and water and the layers separated. The aqueouslayer is extracted with ethyl acetate. The combined organic layers arewashed with brine, dried (Na₂SO₄) and concentrated under reducedpressure to yield 0.11 g (100%) of the pure desired product as a whitesolid MS ESI (+) m/z 482, 484 (M+, Br pattern) detected.

Step C:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-vinyloxy-ethoxy)-amide (11s)

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid 11r (0.11 g, 0.23 mmol) is dissolved in N,N-dimethylformamide (2mL). HOBT (0.037 g, 0.27 mmol) and triethylamine (0.094 mL, 0.68 mmol)are added. Then O-(2-vinyloxy-ethyl)-hydroxylamine (0.028 g, 0.27 mmol)and EDCl (0,056 g, 0.29 mmol) are added and the reaction mixture isstirred at room temperature under N₂ until HPLC shows the reaction iscomplete (2-3 days). The reaction mixture is poured into a separatoryfunnel, diluted with ethyl acetate and water and the layers separated.The ethyl acetate is washed successively with aqueous saturated NH₄Cl(2×), brine (1×), aqueous saturated sodium bicarbonate (2×), water (1×),and brine (1×), dried (Na₂SO₄) and concentrated under reduced pressure.The resulting solid is purified by FCC (eluting with 15:1 methylenechloride:methanol) to yield 0.039 g (79%) of the pure desired product asan off-white solid MS ESI (+) m/z 567, 569 (M+, Br pattern) detected.

Step D:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-hydro-ethoxy)-amide 11p

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-vinyloxy-ethoxy)-amide 11s (0.039 g, 0.068 mmol) is dissolved inethanol (2 mL) and aqueous 2 M HCl (200 uL) is added. The reactionmixture is stirred at room temperature for 30 minutes. The reactionmixture is diluted with water and then neutralized with aqueous 2 M NaOH(˜200 μL) until pH 7 and concentrated under reduced pressure. Theresidue is partitioned between ethyl acetate and brine in a separatoryfunnel and the layers separated. The ethyl acetate layer is dried(Na₂SO₄) and concentrated under reduced pressure to yield 0.034 g (91%)of the pure desired product as an off-white solid. MS ESI (+) m/z 541,543 (M+, Br pattern) detected; ¹H NMR (400 MHz, CD₃OD) δ 8.29 (s, 1H),7.75 (s, 1H), 7.49 (d, 1H), 7.18 (dd, 1H), 6.40 (dd, 1H), 4.40 (dd, A ofABX pattern, 1H), 4.28 (dd, B of ABX pattern, 1H), 3.92 (m, X of ABXpattern, 1H), 3.66 (t, 2H), 3.35 (m, 1H), 1.89 (m, 1H), 1.76 (m, 1H),2.28 (s, 3H), 1.54 (m, 3H), 1.30 (m, 1H). ¹⁹F NMR (376 MHz,CD₃OD)−134.87 (s).

Example 28

The following compounds are prepared by methods similar to thatdescribed in Example 27 by using the appropriate methyl ester andalkylating agent (Step A) and the appropriate hydroxylamine in (Step C).11t

11u

11v

11w

11x

11y

11z

11aa

Example 29

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (11bb)

Step A:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carboxylicacid methyl ester 11cc

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b (1.55 g, 3.89 mmol) is dissolved in 15 mL DMF underN₂, K₂CO₃ (0.70 g, 5.06 mmol) is added followed by methyl vinyl sulfone(0.41 mL, 4.67 mmol). After stirring 16 hours at room temperature, thereaction mixture is diluted with ethyl acetate and water. The layers areseparated and the organic layer is washed with water (3×) and brine. Thecombined aqueous washes are extracted with ethyl acetate. The combinedorganic extract are dried (MgSO₄) and concentrated under reduced.Purification by dissolving the residue in methylene chloride andprecipitating with diethyl ether, repeated several times, gives 1.16 g(50%) pure desired product as a yellow solid: MS APCI (+) m/z 506, 504(M+ Br pattern) and 400, 398 (M− methyl ethyl sulfone Br pattern).

Step B:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11bb

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carboxylicacid methyl ester 11cc is subjected to methods previously described togive6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide: MS APCI (+) m/z 561, 559 (M+, Br pattern)and MS APCI (−) m/z 559, 557 (M− Br pattern) detected; ¹H NMR (400 MHz,DMSO-d₆)

11.75 (s, 1H), 8.47 (s, 1H), 8.04 (s, 1H), 7.77 (s, 1H), 7.62 (d, 1H),7.28 (dd, 1H), 6.40 (dd, 1H), 4.78 (t, 2H), 3.82 (t, 2H), 3.62 (d, 2H),3.07 (s, 3H), 1.02 (m, 1H), 0.49 (m, 2H), 0.21 (m, 2H); ¹⁹F NMR (376MHz, DMSO-d₆)−132.66 (s).

Example 30

The following compounds were prepared similarly using the appropriatemethyl ester and Michael acceptor and methods described previously. 11dd

11ee

11ff

11gg

11hh

11ii

11jj

11kk

11ll

11mm

11nn

Example 31

[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-bromo-2-methyl-phenyl)-amine(24a)

Step A:6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide 20a

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8a (0.051 g, 0.135 mmol) is suspended in EtOH (5 mL)and hydrazine hydrate (0.118 g, 2.023 mmol) is added. The reactionmixture is heated at reflux for 16 hours. The reaction mixture isconcentrated under reduced pressure and purified by FCC eluted with 97:3ethyl acetate:MeOH to give 0.041 g (81%) of clean desired product: LC/MSESI (+)m/z 378, 380 (M+ Br pattern) detected.

Step B:[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-bromo-2-methyl-phenyl)-amine24a

6-(4-Bromo-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide 20a (0.041 g, 0.109 mmol) is suspended in 1,4-dioxane(1.5 mL) and 36 μL of a 3 M solution of cyanogen bromide in methylenechloride is added. NaHCO₃ (9 mg, 0.109 mmol) in water (1.5 mL) is thenadded. After 16 hours, the reaction mixture is diluted with water andbrine and extracted with THF. The combined organic extracts are dried(Na₂SO₄) and concentrated under reduced pressure. Purification by FCCeluted with 98:2 ethyl acetate:MeOH gives 24 mg (55%) of pure desiredproduct as a yellow solid: LC/MS ESI (+) m/z 403, 405 (M+ Br pattern)detected; ¹H-NMR (400 MHz, DMSO-d₆) δ 12.97 (s, 1H), 8.42 (s, 1H), 7.94(s, 1H), 7.74 (s, 1H), 7.36 (s, 2H), 7.33 (d, 1H), 7.15 (d, 1H), 6.40(bs, 1H), 2.34 (s, 3H).

Example 32

[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-chloro-2-methyl-phenyl)-amine(24b)

[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-chloro-2-methyl-phenyl)-amine24b is prepared as described in example 31 staring with6-(4-chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8e. LC/Ms ESI (+) m/z 359, 361 (M+ Cl pattern)detected; ¹H NMR (400 MHz, DMSO-d₆) δ 8.42 (s, 1H), 8.00 (bs, 1H), 7.78(bs, 1H), 7.48 (s, 2H), 7.22 (s, 1H), 7.04 (d, 1H), 6.48 (bs, 1H), 2.37(s, 3H).

Example 33

[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-chloro-2-methyl-phenyl)-amine(24c)

[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-bromo-2-chloro-phenyl)-amine24c is prepared as described in example 31 starting with6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b. NS APCI (+) m/z 425, 423 (M+ Br pattern) and MSAPCI (−) m/z 423, 421 (M− Br pattern) detected.

Example 34

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide (20b)

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide 20b is prepared as described in example 31, step A from6-(4-chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8e. LC/MS ESI (+) m/z 334, 336 (M+ Cl pattern)detected; ¹H NMR (400 MHz, DMSO-d₆) δ 13.09 (bs, 1H), 9.98 (s, 1H), 8.40(s, 1H), 8.17 (bs, 1H), 7.64 (bs, 1H), 7.20 (s, 1H), 7.03 (d, 1H), 6.41(s, 1H), 4.49 (s, 2H), 2.23 (s, 3H).

Example 35

5-[6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-[1,3,4]oxadiazol-2-ol(22a)

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide 20b (0.050 g, 0.150 mmol) is suspended in PhMe (2 mL) anda 20% phosgene solution in PhMe (0.24 mL, 0.45 mmol) is added. Thereaction mixture is stirred at reflux under N₂ for 1 h and then cooledto rt. The reaction mixture is quenched by the addition of 1:1 mixtureof THF and 10% HCl (20 mL). The layers are separated and the aqueouslayer is extracted with THF (3×). The combined organic layer is washedwith brine, dried (Na₂SO₄) and concentrated under reduced pressure togive 54 mg (99%) of desired product as a yellow solid: LC/MS ESI (+) m/z360, 362 (M+ Cl pattern) detected; ¹H NMR (400 MHz, DMSO-d₆) δ 12.64 (s,1H), 8.83 (s, 1H), 7.88 (s, 1H), 7.30 (s, 1H), 7.20 (d, 1H), 7.00 (dd,1H), 6.38 (dd, 1H), 2.30 (s, 3H).

Example 36

(4-Chloro-2-methyl-phenyl)-(4-fluoro-6-[1,3,4]oxadiazol-2-yl-1H-benzoimidazol-5-yl)-amine(21a)

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide 20b (0.048 g, 0.144 mmol) is suspended in 3 mL absoluteEtOH and HC(OEt)₃ (0.60 mL, 3.54 mmol) is added followed by catalyticpTsOH.H₂O. The reaction mixture is heated to reflex under N₂. After 2 h,the reaction mixture is cooled to room temperature and concentratedunder reduced pressure. Purification by flash column chromatography(97.3 ethyl acetate:MeOH) gives 36 mg (73%) desired product as a lightyellow solid. LC/MS ESI (+) m/z 344, 346 (M+ Cl pattern) detected; ¹HNMR (400 MHz, DMSO-d₆)

13.10 (bs, 1H), 9.39 (s, 1H), 8.49 (s, 1H), 8.10 (bs, 1H), 7.78 (bs,1H), 7.20 (d, 1H), 7.00 (dd, 1H), 6.41 (bs, 1H), 2.18 (s, 3H).

Example 37

5-[6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-[1,3,4]oxadiazol-2-thiol(23a)

6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid hydrazide 20b (0.050 g, 0.150 mmol) is suspended in 3 mL absoluteEtOH and cooled to 0° C. under N₂. CS₂is added (26 mg, 0.346 mmol)followed by powdered KOH (8 mg, 0.150 mmol). After stirring at 0° C. for30 minutes, the reaction mixture is heated to reflux. After 3.5 hours,the reaction mixture is quenched by the addition of water, followed bythe addition of ethyl acetate and 1N HCl. The layers are separated andthe aqueous layer is extracted with ethyl acetate. The combined organicextract are dried (Na₂SO₄) and concentrated under reduced pressure togive the desired product as a yellow solid: LC/MS ESI (+) m/z 376, 378(M+ Cl pattern) detected; ¹H NMR (400 MHz, DMSO-d₆)

8.51 (s, 1H), 7.92 (s, 1H), 7.19 (s, 1H), 7.12 (s, 1H), 6.98 (d, 1H),6.29 (d, 1H), 2.28 (s, 3H).

Example 38

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methylamide (11oo)

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid 10c (0.029 g, 0.076 mmol) is dissolved in N,N-dimethylformamide(1.1 mL). HOBT (0.016 g, 0.10 mmol), triethylamine (0.028 mL, 0.20mmol), methylamine (0.059 mL, 0.12 mmol, 2 M solution intetrahydrofuran), and EDCl (0.019 g, 0.10 mmol) are added consecutivelyto the reaction mixture at room temperature. The solution is stirred atroom temperature for 16 hours under N₂. The reaction mixture is pouredinto a separatory funnel and diluted with ethyl acetate and water andthe layers separated. The ethyl acetate layer is washed successivelywith aqueous saturated NH₄Cl (2×), brine (1×), aqueous saturated sodiumbicarbonate (2×), water (1×), and brine (1×), dried (MgSO₄) andconcentrated under reduced pressure. The resulting solid is purified byFCC (eluting with 19:1 methylene chloride:methanol) to yield 0.013 g(42%) of the pure desired product. MS APCI (+) m/z 397, 399 (M+, Brpattern) detected; ¹H NMR (400 MHz, DMSO-d₆) δ 8.76 (broad s, 1H), 8.69(m, 1H), 8.41 (s, 1H), 7.76 (s, 1H), 7.63 (d, 1H), 7.30 (dd, 1H), 6.50(dd, 1H), 2.76 and 2.75 (s and s, 3H total, amide rotamers). ¹⁹F NMR(376 MHz, DMSO-d6)−132.69 (s).

Example 39

The following compounds are prepared using methods similar to thatdescribed above in Example 38 by using the appropriate carboxylic acidand amine. In those cases that contain two amine functionalities, theappropriate mono Boc protected amine is used in the coupling reactionand the Boc group is later removed in a final step under standard TFAdeprotection conditions. 11pp

11qq

11rr

11ss

11tt

11uu

11vv

11ww

11xx

Example 40

[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-methanol(10e)

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid methyl ester 8b (1.06 g, 2.65 mmol) is suspended in tetrahydrofuran(25 mL) and cooled to −78° C. Lithium aluminum hydride (8.03 mL, 8.03mmol, 1 M solution in tetrahydrofuran) is added dropwise to the reactionmixture. After stirring for 10 minutes at −78° C., the reaction mixtureis warmed to 0° C. and becomes a homogeneous solution. The reactionmixture is stirred for 5 minutes at 0° C. and then cooled again to −78°C. The reaction mixture is quenched with MeOH, diluted with Rochelle'ssalt, warmed to room temperature and stirred for 1 hour. The reactionmixture is then poured into a separatory funnel, diluted with ethylacetate, and the layers separated. The aqueous phase is extracted withethyl acetate. The combined ethyl acetate layers are dried (Na₂SO₄) andconcentrated under reduced pressure to yield 0.98 g (100%) of the puredesired product as a pale yellow solid. MS ESI (+) m/z 370, 372 (M+, Brpattern) detected.

Example 41

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde(10f)

[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-yl]-methanol10e (0.96 g, 2.58 mmol) is dissolved in tetrahydrofuran/acetone (1:1, 15mL), and MnO₂ (2.24 g, 25.8 mmol) is added. The reaction mixture isstirred at 50° C. for 10 hours under N₂. The reaction mixture isfiltered through silica gel and eluted with methylene chloride/methanol(10:1, 1 L). The filtrate is concentrated under reduced pressure to asmall volume and then filtered through an Acrodisc syringe filter toremove small amounts of MnO₂ that passed through the silica gel. Thefiltrate is concentrated under reduced pressure and the residue ispurified by flash column chromatography (eluting with 20:1 methylenechloride:methanol) to yield 0.81 g (85%) of the pure desired product asa bright yellow solid. MS ESI (+) m/z 368, 370 (M+, Br pattern)detected.

Example 42

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5-yl]-2-hydroxy-ethanone(10g)

Step A:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5-yl]-2-methoxymethoxy-ethanol10i

To a solution of tributyl-methoxymethoxymethyl-stannane (864 mg, 2.37mmol, prepared by the procedure reported in J. Org. Chem. 1988, 53,4131) in THF (8 mL) at −78° C. is added n-BuLi (0.94 mL, 2.35 mmol, 2.5M solution in hexane). After stirring for 3 minutes, a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbaldehyde10h (59 mg, 0.15 mmol) in THF (2 mL) is added at −78° C. After stirringfor 40 minutes at −78° C., the reaction is quenched with saturatedaqueous NH₄Cl at −78° C., warmed to room temperature, and diluted withEtOAc. The organic layer is washed with brine, dried over MgSO₄,filtered, concentrated, and purified by flash chromotography (1.5% MeOHin CH₂Cl₂) to give the desired product (45 mg, 64%): MS APCI (+) m/z458, 460 (M+, Br pattern) detected.

Step B:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5-yl]-2methoxymethoxy-ethanone10j

A solution of1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-yl]-2-methoxymethoxy-ethanol10i (44 mg, 0.096 mmol) and the Dess-Martin periodinane (49 mg, 0.12mmol) in CH₂Cl₂ (1.5 mL) is stirred for 1.5 hours at rt. The reactionmixture is diluted with ether (3 mL). Saturated aqueous NaHCO₃ (1 mL)containing sodium thiosulfate pentahydrate (74 mg) is added. Theresulting mixture is stirred for 10 minutes and diluted with EtOAc. Theorganic layer is washed with saturated aqueous NaHCO₃ and brine, driedover MgSO₄, filtered, and concentrated in vacuo to give a crude materialwhich is purified by flash chromatography (1.5% MeOH in CH₂Cl₂) toafford the desired product (31 mg, 71%): MS APCI (+) m/z 456, 458 (M+,Br pattern) detected.

Step C:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5-yl]-2-hydroxy-ethanone10g

A mixture of1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-yl]-2methoxymethoxy-ethanone10j (15 mg, 0.033 mmol), 10% aqueous HCl (0.3 mL), methanol (0.01 mL),and water (0.05 mL) is stirred for 3 days at rt. The reaction mixture isneutralized with saturated aqueous NaHCO₃, and diluted with EtOAc. Theorganic layer is washed with brine, dried over MgSO₄, filtered,concentrated in vacuo, and purified by flash chromatography (1.5% MeOHin CH₂Cl₂) to afford the desired product (7.3 mg, 54%): MS APCI (+) m/z412, 414 (M+, Br pattern) detected; ¹H NMR (400 MHz, acetone-d₆) δ 8.64(s, 1H), 8.34 (s, 1H), 8.16 (s, 1H), 7.58 (d, 1H), 7.31 (dd, 1H), 6.59(dd, 1H), 4.94 (s, 2H), 4.06 (s, 3H); ¹⁹F NMR (376 MHz,acetone-d6)−132.45 (s).

Example 43

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-hydroxy-ethanone(10k)

Step A:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-yl]-2-methoxymethoxy-ethanol10l

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde10f is treated with tributyl-methoxymethoxymethyl-stannane according tothe procedure described in Example 42, Step A to yield compound 10l. MSAPCI (+) m/z 444, 446 (M+, Br pattern) detected.

Step B:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methoxymethoxy-ethanone10m

To a solution of oxalyl chloride (0.11 mL, 0.22 mmol) in CH₂Cl₂ (1 mL)at −78° C. is added DMSO (0.016 mL, 0.22 mmol). After stirring for 3minutes, a solution of1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methoxymethoxy-ethanol10l (25 mg, 0.056 mmol) in methylene chloride (1 mL) is added. Theresulting solution is stirred for 30 minutes at −78° C. TEA (0.1 mL,0.71 mmol) is added. The reaction mixture is slowly warmed to roomtemperature, stirred for 5 minutes at room temperature, and diluted withwater and CH₂Cl₂. The organic layer is separated, dried over MgSO₄,filtered, and concentrated to give the crude product which is directlyused without further purification.

Step C:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-hydroxy-ethanone10k

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2methoxymethoxy-ethanone10m is deprotected according to the procedure described in Example 42,Step C to yield compound 10k. MS APCI (+) m/z 398, 400 (M+, Br pattern)detected; ¹H NMR (400 MHz, CD₃OD) δ 8.38 (s, 1H), 8.04 (s, 1H), 7.52 (d,1H), 7.22 (dd, 1H), 6.53 (dd, 1H), 4.90 (m, 2H); ¹⁹F NMR (376 MHz,CD₃OD)−133.96 (s, 1F).

Example 44

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-ethoxy-ethanone(10n)

Step A:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-ethoxy-ethanol10o

To a solution of lithiomethyl ethyl in THF (6 mL) (prepared from4,4′-di-tert-butylbiphenyl (5.85 mg, 2.20 mmol). Li (18 mg, 2.59 mmol),and EtOCH₂Cl (0.20 mL, 2.05 mmol) by the procedure reported inTetrahedron 1996, 52, 1943) is added a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde10f (29 mg, 0.080 mmol) in THF (1 mL) at −78° C. The resulting solutionis stirred for 1 hour and then quenched with saturated aqueous NH₄Cl at−78° C., warmed to room temperature, and extracted with EtOAc. Theorganic layer is washed with brine, dried over MgSO₄, filtered,concentrated in vacuo, and purified by flash chromotography (100% CH₂Cl₂to 3% to 5% MeOH in CH₂ Cl₂) to give the desired product (15 mg, 44%):MS APCI (+) m/z 428, 430 (M+, Br pattern) detected.

Step B:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-ethoxy-ethanone10n

The title compound is prepared from1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-ethoxy-ethanol10o according to the procedure described in Example 42, Step B exceptthat the reaction mixture is not treated with saturated aqueous NaHCO₃containing sodium thiosulfate pentahydrate. MS APCI (+) m/z 426, 428(M+, Br pattern) detected; ¹H NMR (400 MHz, CD₃OD) δ 8.36 (s, 1H), 8.04(s, 1H), 7.51 (d, 1H), 7.21 (dd, 1H), 6.51 (dd, 1H), 4.76 (s, 2H), 3.57(q, 2H), 1.19 (t, 3H); ¹⁹F NMR (376 MHz, CD₃OD)−133.96 (s).

Example 45

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methoxy-ethanone(10p)

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methoxy-ethanone10p is prepared from6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde10f and lithiomethyl methyl ether by the procedures described in Example44. MS APCI (+) m/z 412, 414 (M+, Br pattern) detected.

Example 46

2-Benzyloxy-1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-ethanone(10q)

Step A:2-Benzyloxy-1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-ethanol10r

To a solution of benzyloxymethyl lithium in THF (2 mL, prepared fromn-Bu₃SnCH₂OBn (505 mg, 1.23 mmol) and n-BuLi (0.49 mL, 1.22 mmol, 2.5 Msolution in hexane) by the procedure reported in J. Am. Chem. Soc. 1978,100, 1481) is added a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde10f a (51 mg, 0.14 mmol) in THF (3 mL) at −78° C. The resulting solutionis stirred for 1 hour at −78° C. The reaction is quenched with saturatedaqueous NH₄Cl, and extracted with EtOAc. The organic layer is dried overMgSO₄, filtered, concentrated in vacuo, and purified by flashchromatography (100% CH₂Cl₂ to 3% MeOH in CH₂Cl₂) to afford the desiredproduct (46 mg, 68%): MS APCI (+) m/z 490, 492 (M+, Br pattern)detected. Step B:2-Benzyloxy-1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-ethanone10q

The title is prepared from2-benzyloxy-1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-ethanol10r by the procedure described in Example 42, Step B except that thereaction mixture is not treated with saturated aqueous NaHCO₃ containingsodium thiosulfate pentahydrate: MS APCI (+) m/z 488, 490 (M+, Brpattern) detected; ¹H NMR (400 MHz, CD₃OD) δ 8.37 (s, 1H), 8.02 (s, 1H),7.51 (d, 1H), 7.26 (m, 5H), 7.19 (dd, 1H), 6.46 (dd, 1H), 4.77 (s, 2H),4.58 (s, 2H); ¹⁹F NMR (376 MHz, CD₃OD)−134.52 (s).

Example 47

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methanesulfonyl-ethanone(10s)

Step A:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methanesulfonyl-ethanone10t

To a solution of methyl sulfone (65 mg, 0.68 mmol) in THF (1.5 mL) isadded a solution of n-BuLi (0.27 mL, 0.68 mmol, 2.5 M solution inhexane) at −78° C. After stirring for 5 minutes, HMPA (0.1 mL) is added.After stirring for additional 10 minutes, a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde10f (26 mg, 0.069 mmol) in THF (1 mL) is added. The resulting solutionis stirred for 1.5 hours at −78° C. The reaction is quenched withsaturated aqueous NH₄Cl, warmed to room temperature, and diluted withEtOAc. The organic layer is washed with water, dried over MgSO₄,filtered, concentrated in vacuo, and purified by flash chromatography(3% MeOH in CH₂Cl₂) to give the crude desired product (31 mg, 96%) whichis used directly without further purification: MS APCI (+) m/z 462, 464(M+, Br pattern) detected.

Step B:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methanesulfonyl-ethanone10s

The title compound is prepared from1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-methanesulfonyl-ethanone10t by the procedure described in Example 42, Step B except that thereaction mixture is not treated with saturated aqueous NaHCO₃ containingsodium thiosulfate pentahydrate. MS APCI (+) m/z 460, 462 (M+, Brpattern) detected; ¹H NMR (400 MHz, acetone-d₆) δ 8.44 (s, 1H), 8.33 (s,1H), 7.59 (s, 1H), 7.32 (d, 5H), 6.68 (dd, 1H), 5.00 (s, 1H), 3.15 (s,3H); ¹⁹F NMR (376 MHz, acetone-d₆)−132.97 (s).

Example 48

1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-ethane-1,2-diol(10u)

Step A:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-(isopropoxy-dimethyl-silanyl)-ethanol10v

To a solution of the Grignard reagent prepared from Mg and chloromethyldimethylisopropoxy silane (Org. Synth. 1992, 69, 96) [4.4 mL, 3.26 mmol,0.74 M solution (based on 90% purity)] in THF, is added a solution of6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carbaldehyde10f (200 mg, 0.54 mmol) in THF (1 mL) at −78° C. After stirring for 1hour at −78° C., the reaction is quenched with saturated aqueous NH₄Cl,and extracted with EtOAc. The organic layer is dried over MgSO₄,filtered, concentrated in vacuo to afford the crude desired productwhich is directly used without further purification.

Step B:1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-ethane-1,2-diol10u

To the crude1-[6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-(isopropoxy-dimethyl-silanyl)-ethanol10v in MeOH-THF (5 mL-5 mL) is added KHCO₃ (54 mg, 0.54 mmol), and KF(74 mg, 1.27 mmol), and 30% aqueous H₂O₂ (0.20 mL) at rt. After stirringfor 3.5 hours at room temperature, the reaction mixture is diluted withwater, and extracted with EtOAc. The organic layer is dried over MgSO₄,filtered, concentrated in vacuo, and purified by flash chromatography(8% to 10% MeOH in CH₂Cl₂) to give the desired product (74 mg, 34%): MSAPCI (+) m/z 400, 402 (M+, Br pattern) detected; ¹H NMR (400 MHz, CD₃OD)δ 8.20 (s, 1H), 7.62 (broad s, 1H), 7.47 (d, 1H), 7.14 (dd, 1H), 6.30(d, 1H), 4.96 (t, 1H), 3.64 (m, 2H); ¹⁹F NMR (376 MHz, CD₃OD)−136.87(s).

Example 49

-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5-yl]-pyridin-2-yl-methanol(10w)

To a solution of 2-bromopyridine (0.10 mL, 1.04 mmol) in THF (3 mL) at−78° C. is added n-BuLi (0.39 mL, 0.98 mmol, 2.5 M solution in hexane).After stirring for 10 minutes at −78° C., a solution of6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbaldehyde10h (25 mg, 0.064 mmol) in THF (1 mL) is added. The resulting reactionmixture is stirred for 1.5 hours at −78° C., quenched with saturatedaqueous NH₄Cl, and extracted with EtOAc. The organic layer is dried overMgSO₄, filtered, concentrated in vacuo, and purified by flashchromatography (2.5% MeOH in CH₂Cl₂) to afford the desired product (18mg, 62%): MS APCI (+) m/z 461, 463 (M+, Br pattern) detected; ¹H NMR(400 MHz, CD₃OD) δ 8.31 (d, 1H), 8.16 (s, 1H), 7.65 (m, 3H), 7.38 (d,1H), 7.10 (m, 1H), 7.00 (dd, 1H), 6.11 (dd, 1H), 6.05 (s, 1H), 3.94 (s,3H); ¹⁹F NMR (376 MHz, CD₃OD)−135.79 (s).

Example 50

(4-Bromo-2-chloro-phenyl)-(4-fluoro-6-oxazol-5-yl-1H-benzoimidazol-5-yl-amine(10x)

Step A:[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazol-5-yl]-methanol10y

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carboxylicacid methyl ester 11cc (0.300 g, 0.594 mmol) is suspended in a mixtureof EtOH (6 mL) and THF (4 mL) under N₂. NaBH₄ (0.112 g, 2.97 mmol) isadded. After approximately 4 days stirring, reaction mixture is quenchedby the addition of AcOH until the reaction mixture reaches pH 7. Thereaction mixture is concentrated to dryness under reduced pressure andthe reside partitioned between ethyl acetate and water. The layers areseparated and the organic layer is washed with water (3×), brine, anddried (Na₂SO₄). The organic layer is concentrated under reduced pressureuntil a white precipitate forms which is collected by filtration to give0.225 g (70%) clean desired product: LC/MS ESI (+) m/z 478, 476 (M+ Brpattern) detected.

Step B:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carbaldehyde10z

[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazol-5-yl]-methanol10y (0.050, 0.105 mmol) is dissolved in 1:1 THF:acetone (2 mL) under N₂and MnO₂ (0.046 g, 0.524 mmol) is added. The reaction mixture is stirredat room temperature for 16 hours, and then at 55° C. for 5 hours.Additional MnO₂ (0.046 g, 0.524 mmol) is added and the reaction mixturestirred at 55° C. for 2 hours. The reaction mixture is concentrated todryness and the residue dissolved in 10:1 methylene chloride:MeOH. Thesolution is filtered through a silica gel eluted with 10:1 methylenechloride:MeOH. The resulting filtrate is concentrated under reducedpressure to give 41 mg (82%) desired product as a bright yellow solid.

Step C:(4-Bromo-2-chloro-phenyl)-(4-fluoro-6-oxazol-5-yl-1H-benzoimidazol-5-yl-amine10x

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carbaldehyde10z (0.025 g, 0.053 mmol) is suspended in MeOH (2 mL) and K₂CO₃ (0.015g, 0.105 mmol) is added following by tosylmethyl isocyanide (0.011 g0.058 mmol). The reaction mixture is heated to reflux under N₂ for 16hours. After cooling additional tosylmethyl isocyanide (0.011 g, 0.058mmol) is added and the reaction mixture heated to reflux under N₂ for 16hours. The reaction mixture is cooled to room temperature, concentratedunder reduced pressure and dissolved in ethyl acetate. The organicsolution is washed with water and brine. The combined aqueous washes areextracted with ethyl acetate. The combined organic extracts are dried(Na₂SO₄) and concentrated under reduced pressure. Purification by flashcolumn chromatography eluted with 20:1 methylene chloride:MeOH gives 4mg (18%) desired product 10x and 1 mg (4%)(4-bromo-2-chloro-phenyl)-[4-fluoro-1-(2-methanesulfonyl-ethyl)-6-oxazol-5-yl-1H-benzoimidazol-5-yl]-amine.

(4-Bromo-2-chloro-phenyl)-(4-fluoro-6-oxazol-5-yl-1H-benzoimidazol-5-yl-amine10x. LC/MS ESI (+) m/z 409, 407 (M+ Br pattern) detected; ¹H NMR (400MHz, MeOH-d₄)

8.33 (d, 1H), 8.24 (s, 1H), 7.94 (bs, 1H), 7.51 (d, 1H), 7.33 (s, 1H),7.07 (dd, 1H), 6.14 (dd, 1H).

(4-Bromo-2-chloro-phenyl)-[4-fluoro-1-(2-methanesulfonyl-ethyl)-6-oxazol-5-yl-1H-benzoimidazol-5-yl]-amine.LC/MS ESI (+) m/z 515, 513 (M+, Br pattern) detected; ¹H NMR (400 MHz,MeOH-d₄)

8.39 (s, 1H), 8.28 (s, 1H), 8.03 (s, 1H), 7.52 (d, 1H), 7.37 (s, 1H),7.07 (m, 1H), 6.14 (dd, 1H), 3.83 (t, 2H), 2.99 (s, 3H); 1.18 (t, 2H).

Example 51

(4-Bromo-2-chloro-phenyl)-[4-fluoro-6-(3H-imidazol-4-yl)-1H-benzoimidazol-5-yl]-amine(10aa)

Step A:(4-Bromo-2-chloro-phenyl)-{4-fluoro-1-(2-methanesulfonyl-ethyl)-6-[4-(toluene-4-sulfonyl)-4,5-dihydro-oxazol-5-yl]-1H-benzoimidazol-5-yl}-amine10bb

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(2-methanesulfonyl-ethyl)-3H-benzoimidazole-5-carbaldehyde10z (0.050 g, 0.107 mmol) is suspended in EtOH (0.5 mL) under N₂ andtosylmethyl isocyanide (0.020 g, 0.105 mmol) is added followed bycatalytic NaCN (˜1 mg). After 2 hours, 2 mL THF is added to assist withsolubility. After stirring for 16 hours at room temperature, a secondequivalent of tosylmethyl isocyanide (0.020 g, 0.105 mmol) is added.After 8 hours, the reaction mixture is concentrated under reducedpressure and used as is in the next reaction: LC/MS ESI (+) m/z 671, 669(M+ Br pattern) detected.

Step B:(4-Bromo-2-chloro-phenyl)-[4-fluoro-6-(3H-imidazol-4-yl)-1H-benzoimidazol-5-yl]-amine10aa

(4-Bromo-2-chloro-phenyl)-{4-fluoro-1-(2-methanesulfonyl-ethyl)-6-[4-(toluene-4-sulfonyl)-4,5-dihydro-oxazol-5-yl]-1H-benzoimidazol-5-yl}-amine10bb (0.072 g, 0.107 mmol) is treated with 2.4 mL of a 2.0 M NH₃ in MeOHsolution in a sealed pressure tube. The reaction mixture is then heatedto 90° C. with stirring for 20 hours and furthered stirred at roomtemperature for 3 days. The reaction mixture is transferred to a roundbottom flask and concentrated under reduced pressure. Purification byflash column chromatography, twice, eluted with 10:1 methylenechloride:MeOH, followed by successive trituations with methylenechloride and then diethyl ether gives 3 mg (7%) desired product: LC/MSESI (+) m/z 408, 406 (M+ Br pattern) detected; ¹H NMR (400 MHz, MeOH-d₄)

8.23 (s, 1H), 7.87 (s, 1H), 7.74 (s, 1H), 7.46 (m, 1H), 7.32 (d, 1H),7.05 (m, 1H), 6.20 (dd, 1H).

Example 52

6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide (10cc)

Step A: 3-Chloro-2,4-difluoro-5-nitro-benzoic acid 2a

3-Chloro-2,4-difluoro-benzoic acid 1a (3.00 g, 15.6 mmol) is added to astirred solution of concentrated H₂SO₄ (16 mL) and fuming nitric acid(0.85 mL, 20.3 mmol). After 3 hours a precipitate forms. The yellowslurry is poured onto ice water (100 mL). The aqueous mixture isextracted with diethyl ether (333 ). The organic extracts are dried(Na₂SO₄) and concentrated under reduced pressure to give 3.50 g (95%) ofclean desired product as a pale yellow solid.

Step B: 4-Amino-3-chloro-2-fluoro-5-nitro-benzoic acid 3a

Ammonium hydroxide solution (6.88 g, ˜30% in water, 58.9 mmol) is addedto a solution of 3-chloro-2,4-difluoro-5-nitro-benzoic acid 2a (3.5 g,14.7 mmol) in water (16 mL) at 0° C. with stirring. Upon completion ofthe ammonium hydroxide addition the reaction mixture is warmed to roomtemperature. After 5 hours the reaction mixture is cooled to 0° C. andconcentrated HCl is carefully added until the pH of the reaction mixtureis near zero. The solid is collected by filtration and washed with waterand diethyl ether. The solids are transferred to a round bottom flask asa solution in MeOH and EtOAc and concentrated under reduced pressure togive 2.96 g of a yellow solid. The filtrate is partitioned betweendiethyl ether and water and the organic layer is washed with brine. Thecombined organic extracts are dried (Na₂SO₄) and concentrated underreduced pressure to give 0.65 g of product. Recovered a total of 3.61 g(104%) of pure desired product, that is carried forward without furtherpurification.

Step C: 4-Amino-3-chloro-2-fluoro-5-nitro-benzoic acid methyl ester 4a

To a stirred solution of 4-Amino-3-chloro-2-fluoro-5-nitro-benzoic acid3a (3.61 g, 15.4 mmol) in THF (30 mL) and MeOH (10 mL), TMS diazomethane(9.23 mL, 2.0 M solution in hexanes, 18.5 mmol) is added. Aftercompletion of reaction, the reaction mixture is concentrated via rotaryevaporation with acetic acid in the trap. The recovered oily solid istriturated with diethyl ether to provide 1.51 g of yellow solid. Thefiltrate is concentrated and triturated with diethyl ether to give anadditional 0.69 g of yellow solid. A total of 2.20 g (57%) of puredesired product is recovered.

Step D: 4-Amino-3-chloro-5-nitro-2-phenylamino-benzoic acid methyl ester5c

4-Amino-3-chloro-2-fluoro-5-nitro-benzoic acid methyl ester 4a (2.20 g,8.84 mmol) is suspended in MeOH (9.4 mL) and aniline (3.22 mL, 35.4mmol) is added. The reaction mixture is heated to reflux with stirringunder a nitrogen atmosphere. After 19 hours, the reaction is complete.Distilled water (3.22 mL) is added to the reaction mixture and refluxingis continued for one hour. The reaction mixture is cooled to 0° C. in anice bath for 20 minutes. The reaction mixture is filtered and washedwith 3:10 distilled water/MeOH (65 mL total) and then with MeOH. Thesolid is dissolved with CH₂Cl₂ and concentrated under reduced pressureto give 2.40 g (84%) of pure desired product. MS APCI (−) m/z 320.3(M−1) detected.

Step E: 4,5-Diamino-3-chloro-2-phenylamino-benzoic acid methyl ester 6b

4-Amino-3-chloro-5-nitro-2-phenylamino-benzoic acid methyl ester 5c(0.50 g, 1.55 mmol) is dissolved into 2:1 EtOH/MeOH (15.5 mL). Saturatedaqueous NH₄Cl (15 mL), Zn powder (1.02 g, 15.6 mmol), and THF (10 mL)are added. After stirring for 20 hours, the reaction mixture is dilutedwith CH₂Cl₂/THF and water. The organic layer is washed with water (3×).The combined organic extracts are dried (Na₂SO₄) and concentrated underreduced pressure. The solids are triturated with ether to give 0.32 g(70%) clean desired product.

Step F: 7-Chloro-6-phenylamino-3H-benzoimidazole-5-carboxylic acidmethyl ester 7c

4,5-Diamino-3-chloro-2-phenylamino-benzoic acid methyl ester 6b (0.32 g,1.09 mmol) and formamidine acetate (72 mg, 1.64 mmol) in EtOH (36 mL)are heated, with stirring, to 80° C. After 44 hours, the reactionmixture is cooled to room temperature and diluted with EtOAc and washedwith water (3×), saturated NaHCO₃, and brine. The combined organicextracts are diluted (Na₂SO₄) and concentrated under reduced pressure togive 0.33 g (99%) clean desired product as a solid. MS APCI (+) m/z320.3 (M+1) detected.

Step G: 6-(4-Bromo-phenylamino)-7-chloro-3H-benzoimidazole-5-carboxylicacid methyl ester 8g

7-Chloro-6-phenylamino-3H-benzoimidazole-5-carboxylic acid methyl ester7c (0.327 g, 1.08 mmol) is dissolved into DMF (16 mL) and NBS (0.193 g,1.08 mmol) is added. After one hour, the reaction mixture is quenched bythe addition of saturated aqueous NaHSO₃. The reaction mixture is thenpartitioned between EtOAc/THF and water. The organic layer is washedwith water and brine. The combined organic extracts are dried (Na₂SO₄)and concentrated under reduced pressure. The recovered solid istriturated with ether to give 0.225 g (54%) pure desired product. MS ESI(+) m/z 382, 384 (N+, Br pattern) detected.

Step H:6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3H-benzoimidazole-5-carboxylicacid methyl ester 10dd

6-(4-Bromo-phenylamino)-7-chloro-3H-benzoimidazole-5-carboxylic acidmethyl ester 8g (0.225 g, 0.591 mmol) is dissolved in DMF (2 mL) and NCS(79 mg, 0.591 mmol) is added. After the NCS is in solution concentratedNCl (0.005 mL, 0.059 mmol) is added. After 2 hours, sodium bicarbonate,water and NaHSO₃ are added to the reaction mixture. Solids are filteredand washed with water and ether to give 0.141 g (57%) of clean desiredproduct as a tan solid. MS APCI (−) m/z 414, 416 (M−, Br pattern)detected.

Step I:6-(4-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid methyl ester 10ee

6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3H-benzoimidazole-5-carboxylicacid methyl ester 10dd (0.141 g, 0.34 mmol), potassium carbonate (0.141g, 1.02 mmol), and iodomethane (0.063 mL, 1.02 mmol) are dissolved indimethylformamide (3 mL). After 20 hours, the reaction mixture isdiluted with EtOAc and washed with water (3×), potassium carbonate, andbrine. The organic layer is dried (Na₂SO₄) and concentrated to a brownoil. The N3 and N1 alkylated regioisomers are separated by flashchromatography (EtOAc). The recovery of the N3 alkylated regioisomer is20.4 mg (28%). MS ESI (+) m/z 428, 430 (M+, Br pattern) detected.

Step J:6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid 10ff

6-(4-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid methyl ester 10ee (21 mg, 0.048 mmol) is dissolved into 2:1THF/water (1.2 mL) and NaOH (0.190 mL, 1.0 M aqueous solution, 0.190mmol) is added. After stirring for 4 hours the reaction is diluted withwater and acidified to pH 2 addition of 1.0 M HCl. The mixture is thenextracted with 3:1 EtOAc/THF (3×), dried (Na₂SO₄) and concentrated togive quantitative yield of desired product as a white solid. Ms APCI (+)m/z 414, 416 (M+, Br pattern) detected.

Step K:6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-vinyloxy-ethoxy)-amide 10gg

6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid 10ff (32 mg, 0.077 mmol), O-(2-vinyloxy-ethyl)-hydroxylamine (0.010mL, 0.092 mmol), HOBt (13 mg, 0.093 mmol), triethylamine (0.011 mL,0.077 mmol) and EDCl (19 mg, 0.10 mmol) are dissolved intodimethylformamide (1.0 mL) and allowed to stir under a nitrogenatmosphere at room temperature for 24 hours. The reaction mixture isdiluted with EtOAc, washed with water (3×), 10% potassium carbonate(2×), saturated ammonium chloride, brine, dried (Na₂SO₄), andconcentrated under reduced pressure to give 39 mg of 85% pure material.MS APCI (−) m/z 497, 501 *M−, Br pattern) detected.

Step L:6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide 10cc

Hydrochloric acid (0.78 mL, 1.0 M aqueous solution, 0.78 mmol) is addedto a suspension of6-(4-Bromo-2-chloro-phenylamino)-7-chloro-3-methyl-3H-benzoimidazole-5-carboxylicacid 10gg (2-vinyloxy-ethoxy)-amide (39 mg, 0.078 mmol) in MeOH (1 mL).After one hour, the reaction mixture is neutralized to pH 7 andconcentrated under reduced pressure. The solids are dissolved in EtOAc,washed with brine, dried (Na₂SO₄), and concentrated under reducedpressure. Flash chromatography (20:1 CH₂Cl₂/MeOH) provides 9 mg (23%) ofpure product: MS APCI (+) m/z 473, 475 (M+, Br pattern) detected; ¹H NMR(400 MHz, CDCl₃) δ 8.30 (s, 1H), 8.08 (s, 1H), 7.57 (d, 1H), 7.15 (dd,1H), 6.21 (d, 1H), 3.97 (s, 3H), 3.86 (m, 2H), 3.57 (m, 2H).

Example 53

6-(4-Bromo-2-chloro-phenylamino)-3H-benzoimidazole-5-carboxylic acid(2-hydroxy-ethoxy)-amide (10hh)

The above compound is prepared in an analogous fashion to Example 52except that Step I is eliminated. Ms APCI (−) m/z 457, 461 (M−, Brpattern) detected; ¹H NMR (400 MHz, CD₃OD) δ 8.40 (s, 1H), 7.85 (s, 1H),7.50 (d, 1H), 7.14 (dd, 1H), 6.21 (d, 1H), 3.84 (m, 2H), 3.61 (m, 2H).

Example 54

6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-2-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide (10ii)

Step A: 4,5-Diamino-3-fluoro-2-phenylamino-benzoic acid methyl ester 6c

4-Amino-3-fluoro-5-nitro-2-phenylamino-benzoic acid methyl ester 26a(11.44 g, 37.48 mmol) is suspended in ethanol (400 mL) and ammoniumformate (11.80 g, 187.0 mmol) and 20% Pd(OH)₂/C (10.00 g, 18.79 mmol)are added. The reaction mixture is stirred at 95° C. under N₂ for 30minutes. The reaction mixture is cooled to room temperature and thenfiltered through celite, rinsing with ethanol. The filtrate isconcentrated under reduced pressure to give 9.63 g (93%) of the puredesired product as a purple/red solid. MS ESI (+) m/z 276 (M+1)detected.

Step B: 7-Fluoro-2-methyl-6-phenylamino-3H-benzoimidazole-5-carboxylicacid methyl ester 31a

4,5-Diamino-3-fluoro-2-phenylamino-benzoic acid methyl ester 6c (0.20 g,0.73 mmol) is suspended in ethanol (3 mL) and 5 M aqueous NCl (1 mL,5.00 mmol) is added. The reaction mixture is brought to reflux under N₂and then 2,4-pentanedione (0.150 mL, 1.45 mmol) is added. The reactionmixture is stirred at reflux for 60 minutes. The reaction mixture iscooled to room temperature and treated with saturated aqueous NaHCO₃until the pH of the reaction mixture is pH 7 and is then concentratedunder reduced pressure to dryness. The residue is diluted with ethylacetate and water, poured into a separatory funnel and the layerseparated. The ethyl layer is washed with brine, dried (Na₂SO₄) andconcentrated under reduced pressure. The red solid residue is trituratedwith diethyl ether to yield a light brown solid and red filtrate. Thesolid is collected and washed with diethyl ether to yield 0.20 g (91%)of the pure desired product as a light brown solid. MS ESI (+) m/z 300(M+1) detected.

Step C:6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-2-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide 10ii

7-Fluoro-2-methyl-6-phenylamino-3H-benzoimidazole-5-carboxylic acidmethyl ester 31a is converted by the bromination, chlorination,hydrolysis, coupling, and hydrolysis procedures already described toyield the pure desired product as an off-white solid. MS ESI (+) m/z457, 459 (M−, Br pattern) detected; ¹H NMR (400 MHz, CD₃OD) δ 7.58 (s,1H), 7.49 (d, 1H), 7.18 (dd, 1H), 6.41 (m, 1H), 3.91 (t, 2H), 3.65 (t,2H), 2.61 (s, 3H); ¹⁹F NMR (376 MHz, CD₃OD)−135.84 (s).

Example 55

6-(4-Cyano-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide (11yy)

Step A:7-Fluoro-6-(4-iodo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid methyl ester 10jj

7-Fluoro-6-o-tolylamino-1H-benzoimidazole-5-carboxylic acid methyl ester7a (1.47 g, 4.92 mmol) is suspended in 1:1 THF:MeOH mixture (44 mL) andcooled to −78° C. under a nitrogen atmosphere. A solution of NIS (1.66g, 7.39 mmol) in THF (2 mL) is added followed by a MeOH (2 mL) solutionof TsOH.H₂O (1.87 g, 9.84 mmol). After 30 minutes, the reaction mixtureis warmed to 0° C. and 1 mL methylene chloride is added. The reaction isslowly allowed to warm to room temperature with stirring over 16 hours.The reaction mixture is quenched by the addition of 10% Na₂S₂O₄solution. The reaction mixture is then diluted with water and ethylacetate and the layers separated. The aqueous layer is extracted withethyl acetate. The combined organic extracts are dried (Na₂SO₄) andconcentrated under reduced pressure. The recovered solid is trituratedwith MeOH to give 1.45 g (69%) pure desired product: MS ESI (+) m/z 426(M+1) detected; Ms ESI (−) m/z 424 (M−1) detected.

Step B:7-Fluoro-6-(4-iodo-2-methyl-phenylamino)-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid methyl ester 10kk

7-Fluoro-6-(4-iodo-2-methyl-phenylamino)-1H-benzoimidazole-5-carboxylicacid methyl ester 10jj (0.200 g, 0.470 mmol) is suspended in DMF (2 mL)under N₂ and cooled to 0° C. in an ice-water bath. NaH (60% dispersionin oil, 0.018 g, 0.470 mmol) is added. After 10 minutes, the reactionmixture is warmed to room temperature and stirred for 30 minutes. Aftercooling to 0° C., SEMCl (0.087 mL, 0.494 mmol) is added and the reactionis allowed to warm to room temperature with stirring overnight. Thereaction mixture is quenched by the addition of water and brine. Thereaction mixture is extracted with ethyl acetate. The combined organicextracts are washed with water and brine, and dried (MgSO₄) andconcentrated under reduced pressure. Purification by flash columnchromatography eluted with 1:1 hexanes:ethyl acetate gives 0.182 g (70%)of desired product as a 1:1 mixture of N1 and N3 isomers as a whitefoam.

Step C:6-(4-Cyano-2-methyl-phenylamino)-7-fluoro-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid methyl ester 10ll

To a stirred solution of a 1:1 mixture of N1:N3 isomers of7-Fluoro-6-(4-iodo-2-methyl-phenylamino)-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid methyl ester 10jj (0.060 g, 0.108 mmol) in 1 mL DMF at roomtemperature under N₂ is added dppf (2 mg, 0.004 mmol) followed byPd₂dba₃ (2 mg, 0.002 mmol) and Zn(CN)₂ (8 mg, 0.065 mmol) (TetrahedronLett. 1999, 40, 8193-8195). The reaction mixture is heated to 120° C.for 45 minutes. The reaction mixture is cooled to room temperature andquenched by the addition of 5 mL of a 4:1:5 mixture of satNH₄Cl:concentrated NH₄OH:water. The aqueous layer is extracted withethyl acetate. The combined organic extracts are washed with water (3×),brine, and dried (MgSO₄) and concentrated under reduced pressure.Purification by flash column chromatography eluted with 1:1hexanes:ethyl acetate gives 38 mg (77%) of desired product as a 1:1mixture of N1 and N3 isomers: APCI MS (+) m/z 455 (M+1) detected.

Step D:6-(4-Cyano-2-methyl-phenylamino)-7-fluoro-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid 10mm

A 1:1 mixture of N1:N3 isomers of6-(4-cyano-2-methyl-phenylamino)-7-fluoro-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid methyl ester 10ll (31 mg, 0.068 mmol) is hydrolyzed with aqueoussodium hydroxide as described previously to give 26 mg (87%) of desiredproduct.

Step E:6-(4-Cyano-2-methyl-phenylamino)-7-fluoro-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11zz

A 1:1 mixture of N1:N3 isomers of6-(4-cyano-2-methyl-phenylamino)-7-fluoro-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid 10mm (26 mg, 0.059 mmol) is coupled with EDCl and cyclopropylmethyl hydroxylamine hydrochloride as described previously to give 28 mg(93) of desired product: APCI MS (+) m/z 510 (M+1) detected.

Step F:6-(4-Cyano-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11yy

To a slurry of a 1:1 mixture of N1:N3 isomers6-(4-cyano-2-methyl-phenylamino)-7-fluoro-(2-trimethylsilanyl-ethoxymethyl)-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11zz (28 mg, 0.055 mmol) in 0.5 mL EtOH isadded 0.5 mL 10% HCl. The reaction mixture is heated to 50° C. withstirring overnight (Written et al., JOC 1986, 51, 1891-1894). Anadditional 0.5 mL 10% HCl is added and the reaction mixture stirred at70° C. overnight. The reaction mixture is cooled to room temperature andneutralized to pH ˜8 with 1.5 mL 1N NaOH. The reaction mixture isextracted with ethyl acetate, dried (MgSO₄) and concentrated underreduced pressure to give 14 mg (60%) of 90% pure product as a mixture ofrotatomers: MS APCI (+) m/z 380 (M+1) detected; MS APCI (−) m/z 378(M−1) detected; ¹H NMR (400 MHz, MeOH-d₄) δ 8.41 (bs, 1H), 7.75 (m, 1H),7.50 (s, 1H), 7.38 (d, 1H), 6.51 (m, 1H), 3.72 (d, 0.5H), 3.65 (d,1.5H); 2.41 (s, 3H), 0.98 (1H, m), 0.58 (d, 1.5H), 0.40 (d, 0.5H), 0.25(d, 1.5H), 0.19 (d, 0.5H).

Example 56

6-(4-Ethynyl-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11aaa

Step A:7-Fluoro-6-(2-methyl-4-trimethylsilanylethynyl-phenylamino)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide 11bbb

7-Fluoro-6-(4-iodo-2-methyl-phenylamino)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxyamide 11ccc (0.025 g, 0.052 mmol) is dissolved in1:1 acetonitrile/triethylamine (0.50 mL). Ethynyltrimethylsilane (0.013mL, 0.092 mmol), Pd(PPh₃)₂Cl₂ (0.004 g, 0.006 mmol), and CuI (0.002 g,0.011 mmol) are added consecutively and the reaction mixture is stirredat 60° C. for 1 hour under N₂. The reaction mixture is cooled to roomtemperature and concentrated under reduced pressure. The residue ispurified by FCC (eluting with 20:1 methylene chloride:methanol) to yield0.020 g (87%) of the desired product.

Step B:6-(4-Ethynyl-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxyamide 11aaa

7-Fluoro-6-(2-methyl-4-trimethylsilanylethynyl-phenylamino)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxyamide 11bbb (0.020 g, 0.044 mmol) is dissolved intetrahydrofuran (0.50 mL) and the reaction solution is cooled to 0° C.TBAF (50 μL, 0.050 mmol, 1 M solution in tetrahydrofuran) is added. Thereaction mixture is warmed to room temperature and additional TBAF (25μL, 0.025 mmol, 1 M solution in tetrahydrofuran) is added. The reactionis stirred at 50° C. for 2 hours under N₂. The reaction mixture iscooled to room temperature, a few drops of H₂O are added and then it isconcentrated under reduced pressure. The residue is purified by FCC(eluting with 20:1 methylene chloride:methanol) to yield 0.011 g (65%)of the pure product. MS APCI (−) m/z 377 (M−1) detected; ¹H NMR (400MHz, CDCl₃) δ 10.56 (broad s, 1H), 8.12 (s, 1H), 7.99 (s, 1H), 7.28 (s,1H), 7.11 (d, 1H), 6.42 (broad, 1H), 3.70 (br s, 2H), 2.96 (d, 1H), 2.37(s, 3H), 0.85 (m, 1H), 0.50 (m, 2H), 0.22 (m, 2H).

Example 57

Phosphoric acidmono-(2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethyl)ester (29nnn)

3.000 g of6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide, 660 mg of tetrazole and 2.80 mL ofdi-tertbutyl diisopropyl-phosphoramidite were dissolved/suspended in 30mL of anhydrous DMF under an atmosphere of dry N₂. The reaction mixturewas stirred for about 3 hrs after which time the reaction was cooled to−78° C. and 2.50 mL of 70% tert-butyl hydrogen peroxide was added. Thecooling bath was then taken away and the reaction was slowly warmed upto room temperature and reacted over night. The reaction was thenpartitioned between a solution of ethyl ether/ethyl acetate (5:1) andsaturated aqueous NaHCO₃. The organic layer was saved and successivelywashed with 10% aqueous sodium sulfite, 3 times with water and finallywith brine. The resulting organic layer was dried over MgSO₄, filteredand concentrated under vacuum. The residue was dissolved in 45 mL of asolution for about 2 hours after which time it was concentrated undervacuum and the resulting residue was stirred in methanol for about 1hours. The off-white solid was collected via suction filtration, washedwith methanol followed by ethyl ether and then air-dried to give 2.717 gof the desired product. ¹H NMR (400 MHz, DMSO-d₆) δ11.94 (brs, 1H), 8.40(s, 1H), 7.96 (brs, 1H), 7.75 (s, 1H), 7.60 (s, 1H), 7.25 (dd, 1H), 6.37(dd, 1H), 4.02 (brs, 4H); 19F NMR (376 MHz, DMSO-d₆)−132.9 (s, 1F).

Example 58

2-Amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylesterhydrochloric acid salt (29ooo)

Step A: Preparation of 2-tert-butoxycarbonylamino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylester:

Dimethylacetamide (30 mL) was added to a mixture of6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxyethoxy)-amide (3 g, 6.555 mmol), BOC-L-valine (1.710 g,7.866 mmol), HOBT.H₂O (1.004 g, 6.555 mmol) and, TEA (0.929 g, 9.177mmol) in a 500 mL RBF with a magnetic stirrer and nitrogen line. After 5minutes, EDCl (1.759 g, 9.177 mmol) saw added to the brown solution inthe 500 mL RBF. The mixture was stirred overnight at room temperature.The reaction mixture was added slowly to 210 mL of deionozied water withmechanical stirring. The white slurry was stirred for 1 hour, filteredthrough a medium fritted funnel. The cake was washed with about 100 mLof water and pulled air through the cake for 10-15 minutes. Tert-butylmethyl ether (70 mL) saw added to the cake and stirred with a spatula.The vacuum filtration was continued. The solids were dried under vacuumat 50° C. to give 2 g of the product containing about 80% of theundesired isomer. The t-butyl methyl ether layer was concentrated to afoam (2.18 g) enriched in the desired product comprising a mixture ofisomers.

Step B: Preparation of 2-amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylesterhydrochloric acid salt:

A mixture of 2-tert-butoxycarbonylamino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylesterand its N-acylated isomer (0.6 g) were treated with 1.5 mL oftrifluoroacetic acid for 0.5 hours. An HPLC trace of the reactionmixture showed the reaction was complete. The mixture was concentratedon a rotary evaporator and pumped on high vacuum to give yellow oil (0.9g). The deprotected samples were submitted for HPLC purification to givepurified product as a trifluoroacetic acid salt.

Step C: Preparation of 2-amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylester:

2-amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylestertrifluoroacetic acid salt (474 mg) was suspended in 80 mL of EtOAc.Addition of 15 mL of saturated aqueous NaHCO₃ solution gave a biphasicmixture and dissolved most of the solids. Water (5 mL) was added todissolve some turbidity seen in the aqueous layer. The layers wereseparated and the aqueous layer was extracted with 20 mL of EtOAc. Thecombined EtOAc layer was diluted with 50 mL of t-butyl methyl ether andwashed with 2×25 mL of water, 10 mL of brine, dried with Na₂SO₄ andfiltered. The organic layer was concentrated to a foam, concentratedwith 2×10 mL of t-butyl methyl ether and dried under vacuum to give 340mg (86%) of the free base as a white solid. ¹H NMR (400 MHz, DMSO-d₆ andD₂O exchange) δ: 0.82 (d, J=6.7 Hz, 3H), 0.87 (d, J=6.7 Hz, 3H), 1.86(m, 1H), 3.14 (d, J=5.0 Hz, 1H), 3.90 (s, 3H), 4.03 (m br, 2H), 4.24 (dbr, 2H), 6.36 (dd, J_(H-H)=8.8 Hz, J_(F-H)=4.4 Hz virtual coupling, 1H);7.26 (dd, J=8.8 Hz, 2.3 Hz, 1H), 7.60 (d, J=2.26 Hz, 1H), 7.75 (s, 1H),8.40 (s, 1H), ¹⁹F NMR (376 MHz, DMSO-d₆ and D₂O exchange) δ: −133.03.

Step D: Preparation of 2-Amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylesterhydrochloric acid salt:

2-Amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylester(325 mg) was dissolved in about 1.5 mL of absolute EtOH, and 0.35 mL of2N HCl in ether was added to give a thick precipitate. As additional 1.5mL of ether were added to make the slurry thin. The slurry was mixed for5-10 minutes and concentrated on a rotary evaporator. The damp solidswere slightly yellowish and dried under high vacuum at room temperatureovernight to give 0.325 g (94%) of the HCl salt. ¹H NMR (400 MHz,DMSO-d₆) δ: 0.97 (d, J=6.6 Hz, 3H), 1.00 (d, J=6.6 Hz, 3H), 2.20 (m br,1H), 3.91 (s, 3H), 3.95 (s br, 1H), 4.09 (m br, 2H), 4.34 (m br, 1H),4.43 (m br, 1H), 6.37 (dd, J_(H-H)=8.8 Hz, J_(F-H)=4.3 Hz virtualcoupling, 1H), 7.27 (dd, J=8.8 Hz, 2.3 Hz, 1H), 7.65 (d, J=2.2 Hz, 1H),7.78 (s, 1H), 7.92 (s, 1H), 8.37 (s br, 3H), 8.43 (s, 1H), 11.86 (s br,1H). ¹⁹F NMR (376 MHz, DMSO-d₆) δ: −132.98.

Example 59

(S)-Phosphoric acidmono-(2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-(tetrahydropyran-2-ylmethyl)-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethyl)ester (29ppp)

401.2 mg of(S)-6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-(tetrahydropyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-hydroxyethoxy)-amide, 78.6 mg of tetrazole and 320 μL ofdi-tertbutyl diisopropyl-phosphoramidite were dissolved/suspended in 3mL of anhydrous DMF under an atmosphere of dry N₂. The reaction mixturewas stirred for about 4 hrs after which time the reaction was cooled to0° C. and 300 μL of 70% tert-butyl hydrogen peroxide were added. Thecooling bath was then taken away and the reaction was slowly warmed upto room temperature and reacted for 2.5 hrs. The reaction was thenquenched with 10 mL of saturated aqueous sodium thiosulfate. Theresulting solution was partitioned between a solution of ethylether/ethyl acetate (10:1) and saturated aqueous NaHCO₃. The organiclayer was saved and successively washed with water (3×) and then withbrine. The resulting organic layer was dried over MgSO₄, filtered andconcentrated under vacuum. The resulting residue was purified usingsilica gel chromatography eluting with 2:1 Ethyl acetate/dichloromethaneto give 295.3 mg of phosphoric acidmono-(2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-(tetrahydropyran-2-ylmethyl)-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethyl)ester di-tert-butyl ester which was then dissolved in a solution of 5 mLof TFA and 3 mL of dichloromethane under an atmosphere of dry N₂. Thereaction was stirred at room temperature for about 2 hours after whichtime it was concentrated under vacuum. The resulting residue was takenup in methanol and re-concentrated (2×). The resulting residue wasstirred in methanol for about 1 hour. The white solid was collected viasuction filtration, washed with methanol followed by ethyl ether andthen air-dried to give 188.6 mg of the desired product. ¹H NMR (400 MHz,DMSO-d₆) δ 11.83 (brs, 1H), 8.35 (s, 1H), 7.95 (brs, 1H), 7.79 (s, 1H),7.61 (d, 1H), 7.26 (dd, 1H), 6.40 (dd, 1H), 4.37 (m, 1H), 4.26 (m, 1H),4.03 (brs, 4H), 3.85 (m, 1H), 3.70 (m, 1H), 3.27 (m, 1H), 1.80 (m, 1H),1.70 (m, 1H), 1.46 (m, 3H), 1.22 (m, 1H); ¹⁹F NMR (376 MHz,DMSO-d₆)−132.9 (s, 1F).

The invention and the manner and process of making and using it, are nowdescribed in such full, clear, concise and exact terms as to enable anyperson skilled in the art to which it pertains, to make and use thesame. It is to be understood that the foregoing describes preferredembodiments of the present invention and that modifications may be madetherein without departing from the spirit or scope of the presentinvention as set forth in the claims. To particularly point out anddistinctly claim the subject matter regarded as invention, the followingclaims conclude this specification.

The words “comprise”, “comprising,” “include,” “including,” and“includes” when used in this specification and in the following claimsare intended to specify the presence of one or more stated features,integers, components, or steps, but they do not preclude the presence oraddition of one or more other features, integers, components, steps, orgroups thereof. Furthermore, since a number of modifications and changeswill readily occur to those skilled in the art, it is not desired tolimit the invention to the exact construction and process describedabove. Accordingly, all suitable modifications and equivalents may beresorted to falling within the scope of the invention as defined by theclaims that follow.

1. An amino acid prodrug of a compound having the formula

and pharmaceutically accepted salts, prodrugs and solvates thereof,wherein:

is an optional bond, provided that one and only one nitrogen of the ringis double-bonded; R¹, R⁹ and R¹⁰ are independently selected fromhydrogen, halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —OR³, —C(O)R³, —C(O)OR³, NR⁴C(O)OR⁶, —OC(O)R³,—NR⁴SO₂R⁶, —SO₂NR³R⁴, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴,—NR⁵C(NCN)NR³R⁴, —NR³R⁴, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl,C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl),—S(O)_(j)(CR⁴R⁵)_(m)-aryl, aryl, arylalkyl, heteoraryl, heteroarylalkyl,heterocyclyl, heterocyclalkyl, —O(CR⁴R⁵)_(m)-aryl, —NR⁴(CR⁴R⁵)_(m)-aryl,—O(CR⁴R⁵)_(m)-heteroaryl, —NR⁴(CR⁴R⁵)_(m)-heteroaryl,—O(CR⁴R⁵)_(m)-heterocyclyl or —NR⁴(CR⁴R⁵)_(m)-heterocyclyl, where eachalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclylportion is optionally substituted with one to five groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; R³ is selectedfrom hydrogen, trifluoromethyl, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, where eachalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclylportion optionally substituted with one to five groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, phosphate, amino acid residue,polypeptide, hemisuccinate, phosphate esters, dimethylaminoacetate,phosphoryloxymethyloxycarbonyl, phosphonamide, —NR′SO₂R″″, —SO₂NR′R″,—C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″, —SR′,—S(O)R″″, —SO₂R″″, —NR′R″, —NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl, or R³ and R⁴ together with the atom to which they areattached form a 4 to 10 membered carbocyclic, heteroaryl or heterocyclicring, each of which is optionally substituted with one to three groupsindependently selected from halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR′SO₂R″″, —SO₂NR′R″,—C(O)R′, —C(O)OR′, —OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″,—SO₂R″″, —NR′R″, —NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl; R′, R″ and R′″ independently are selected fromhydrogen, lower alkyl, lower alkenyl, aryl and arylalkyl; R″″ isselected from alkyl, lower alkenyl, aryl and arylalkyl; or any two ofR′, R″, R′″ or R″″ together with the atom to which they are attachedform a 4 to 10 membered carbocyclic, heteroaryl or heterocyclic ring,each of which is optionally substituted with one to three groupsindependently selected from halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; R⁴ and R⁵independently represent hydrogen or C₁-C₆ alkyl, or R⁴ and R⁵ togetherwith the atom to which they are attached form a 4 to 10 memberedcarbocyclic, heteroaryl or heterocyclic ring, each of which isoptionally substituted with one to three groups independently selectedfrom halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R″,—NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; R⁶ is selectedfrom trifluoromethyl, C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, where eachalkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl portion isoptionally substituted with one to five groups independently selectedfrom oxo, halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR′SO₂R″″, —SO₂NR′R″, —C(O)R′, —C(O)OR′,—OC(O)R′, —NR′C(O)OR″″, —NR′C(O)R″, —C(O)NR′R″, —SO₂R″″, —NR′R′,—NR′C(O)NR″R′″, —NR′C(NCN)NR″R′″, —OR′, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; R⁷ is selectedfrom hydrogen, C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀cycloalkyl, C₃-C₁₀ cycloalkylalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclyl, heterocyclylalkyl, where each alkyl,alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclyl portionis optionally substituted with one to five groups independently selectedfrom oxo, halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³,—OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —SO₂R⁶, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; W is selected fromheteroaryl, heterocyclyl, —C(O)OR³, —C(O)NR³R⁴, —C(O)NR⁴OR³,—C(O)(C₃-C₁₀ cycloalkyl), —C(O)(C₁-C₁₀ alkyl), —C(O)(aryl),—C(O)(heteroaryl) and —C(O)(heterocyclyl), each of which is optionallysubstituted with 1-5 groups independently selected from —NR³R⁴, —OR³,—R², or C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl, each of whichis optionally substituted with 1 or 2 groups independently selected from—NR³R⁴ and —OR³; R² is hydrogen, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —OR³, —C(O)R³, —C(O)OR³,NR⁴C(O)OR⁶, —OC(O)R³, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —NR⁴C(O)R³, —C(O)NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —NR³R⁴, or C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl,C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl,—S(O)_(j)(C₁-C₆ alkyl), —S(O)_(j)(CR⁴R⁵)_(m)-aryl, aryl, arylalkyl,heteoraryl, heteroarylalkyl, heterocyclyl, heterocyclalkyl,—O(CR⁴R⁵)_(m)-aryl, —NR⁴(CR⁴R⁵)_(m)-aryl, —O(CR⁴R⁵)_(m)-heteroaryl,—NR⁴(CR⁴R⁵)_(m)-heteroaryl, —O(CR⁴R⁵)_(m)-heterocyclyl or—NR⁴(CR⁴R⁵)_(m)-heterocyclyl, where each alkyl, alkenyl, alkynyl,cycloalkyl, aryl, heteroaryl and heterocyclyl portion is optionallysubstituted with one to five groups independently selected from oxo,halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³, —C(O)OR³,—OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴,—NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl, heteroarylalkyl,heterocyclyl, and heterocyclylalkyl; R⁸ is selected from hydrogen,—SCF₃, —Cl, —Br, —F, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, —OR³, —C(O)R³, —C(O)OR³, —NR⁴C(O)OR⁶, —OC(O)R³,—NR⁴SO₂R⁶, —SO₂NR³R⁴, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR⁵C(O)NR³R⁴, —NR³R⁴,C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀cycloalkylalkyl, —S(O)_(j)(C₁-C₆ alkyl), —S(O)_(j)(CR⁴C⁵)_(m)-aryl,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl, —O(CR⁴R⁵)_(m)-aryl, —NR₄(CR⁴R⁵)_(m)-aryl,—O(CR⁴R⁵)_(m)-heteroaryl, —NR⁴(CR⁴R⁵)_(m)-heteroaryl,—O(CR⁴R⁵)_(m)-heterocyclyl or —NR⁴(CR⁴R⁵)_(m)-heterocyclyl, where eachalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl and heterocyclylportion is optionally substituted with one to five groups independentlyselected from oxo, halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶, —SO₂NR³R⁴, —C(O)R³,—C(O)OR³, —OC(O)R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³, —C(O)NR³R⁴, —NR³R⁴,—NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; m is 0, 1, 2, 3, 4or 5; and j is 1 or
 2. 2. A compound according to claim 1 where W is


3. A compound according to claim 1 having the formula


4. A compound according to claim 3 wherein R⁷ is C₁-C₁₀ alkyl, C₃-C₇cycloalkyl, C₃-C₇ cycloalkylalkyl, C₃-C₇ heterocycloalkyl or C₃-C₇heterocycloalkylalkyl, each of which can be optionally substituted with1-3 groups independently selected from oxo, halogen, cyano, nitro,trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, —NR⁴SO₂R⁶,—SO₂NR³R⁴, —C(O)R³, —C(O)OR³, —OC(O)R³, —SO₂R³, —NR⁴C(O)OR⁶, —NR⁴C(O)R³,—C(O)NR³R⁴, —NR³R⁴, —NR⁵C(O)NR³R⁴, —NR⁵C(NCN)NR³R⁴, —OR³, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl; R⁹ is hydrogen or halogen; R¹ is lower alkyl orhalogen; and R⁸ is —OCF₃, Br or Cl.
 5. A compound according to claim 4wherein R⁹ is fluoro.
 6. A compound according to claim 5 wherein R¹ ismethyl or chloro.
 7. A compound according to claim 6 wherein R⁸ ischloro or bromo.
 8. An amino acid prodrug of a compound selected from7-Fluoro-6-(4-bromo-2-methyl-phenylamino)-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide,6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid cyclopropylmethoxy-amide,6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide,6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2,3-dihydroxy-propoxy)-amide,6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-pyran-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide,[6-(5-Amino-[1,3,4]oxadiazol-2-yl)-4-fluoro-1H-benzoimidazol-5-yl]-(4-bromo-2-methyl-phenyl)-amine,1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazol-5-yl]-2-hydroxy-ethanone,1-[6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3H-benzoimidazol-5-yl]-2-hydroxy-ethanone,6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-1,1-dimethyl-ethoxy)-amide,6-(4-Bromo-2-chloro-phenylamino)-7-fluoro-3-(tetrahydro-furan-2-ylmethyl)-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide,6-(4-Chloro-2-methyl-phenylamino)-7-fluoro-3H-benzoimidazole-5-carboxylicacid tert-butoxy-amide,6-(4-Bromo-2-fluoro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide and6-(2,4-Dichloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carboxylicacid (2-hydroxy-ethoxy)-amide.
 9. The compound of claim 8, which is2-amino-3-methylbutyric acid2-{[6-(4-bromo-2-chlorophenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-carbonyl]-aminooxy}-ethylester10. A composition comprising a compound of claim 1 and apharmaceutically acceptable carrier.